INTRODUCTION:

Chitin, a glycopolymer of N-acetylglucosamine which is linked with β-1,4, glycosidic bond, is the second most abundant polysaccharide in nature next to cellulose. Chitin is abundantly present in the cell wall of fungi, exoskeleton of insects and crustacean cells. Marine environment also has a plenty of chitin sources. Chitin exists in nature as crystalline structure in three different forms: α-chitin, β-chitin and γ-chitin^1,2,3 Mohammed et al have studied the extraction of chitin from prawn shell and its conversion into low molecular mass chitosan⁴. As chitin is an insoluble linear polymer, the degradation of chitin can be done by using the chitinolytic enzyme chitinase.

The chitin hydrolyzing enzyme, chitinase is present in a wide range of organisms like virus, fungi, insects, algae, bacteria, animals and some higher plants³. Most of the filamentous fungi produce different kinds of chitinase¹.

Chitinase (EC3.2.1.14) is ubiquitous chitin fragmenting glycosyl hydrolase which hydrolyses the β-1,4-glycosidic bond between the N-acetylglucosamine residue in chitin².

Chitinase is divided into two major categories namely endochitinase and exochitinase. At the same time the degradation of chitin can be done by using the enzyme system: endochitinase, which randomly cleaves the β-1,4-glycosidic bonds of chitin at internal sites in a non-processive mode; exochitinase, which catalyses the release of N,N'-diacetylchitobiose (GlcNAc)₂ from the non-reducing or reducing end of the chitin chain, mostly in a processive mode; N-acetylglucosaminase, which catalyses the conversion of N,N'-diacetylchitobiose (GlcNAc)₂ into beta-1,4-linked N-acetylglucosamine (GlcNAc); and polysaccharide monooxygenase, previously termed chitin-binding protein, which is responsible for destroying the crystalline structure of chitin².

The chitinolytic enzymes are classified into three major families based on their amino acid sequence similarity. Family 18, 19 and 20. Family 18 consist of chitinases from bacteria, fungi, viruses, animals etc. and family 19 contains some plant based chitinase. Family 20 is included in human based chitinase³.

The role of chitinase in the source organisms are different. In vertebrates, it is a part of the digestive tract ⁵. In plants, chitinase is used for the resistance against harmful disease causing pathogen. In fungi and bacteria, the chitinolytic enzyme plays nutritional, morphogenetic and parasitic roles ³.

Several endophytic fungi and bacterium also produce different chitinase enzymes. Quencine MC⁶ and team described the chitinolytic activity of endophytic Streptomyces and the potential for controlling phytopathogenic fungi. They have demonstrated a genetic correlation between chitinase production and pathogen inhibition. Venkatachalam⁷ and team isolated endophytic fungi from different seaweeds and seagrasses for the production of chitinase enzyme and found that Penicillium sp. and a Cladosporium sp. showed higher chitinase activity. Here, the enzyme chitinase was described based on its application towards cosmetics, agriculture and pharmaceutical industries.

There are several applications for the chitinolytic enzymes in the field of biotechnology, medicine, agriculture, bioremediation and various other industries.

Chitinase is an interesting target enzyme for protein engineering¹. Chitinase is used in the protoplast isolation from fungi. It is a potential biocontrol agent against plant pathogen. The genetically engineered chitinase helps in developing fungal resistant transgenic plants and crops 3. Chitinase is involved in the therapeutic regimen of Gaucher disease, asthma, cancer and various other diseases. It is an economical pesticide marker for monitoring the disease progression in plants^2,3. S. Meena et al have studied the effect of metal ions and chemical compounds on chitinase produced by a newly isolated thermotolerant Paenibacillus sp. BISR-047 and its shelf-life⁸.

Chitinase plays a major role in the production of single cell protein, estimation of fungal biomass etc. Another important application of chitinase is it is used for the control of mosquito and morphogenesis³.

Chitinases are involved in the human health care. It is used to produce antifungal drug as it has the antifungal property^3,9. Chitinase plays an important role in asthma and allergic reactions. Chitooligosaccharides has a potential application in pharmacology due to its anti-tumor activity and wound healing property. It also has the anti-inflammatory effect³.

Chitinase is used in treating waste water from many industries^1,5. Chitinase plays a major role in the food industry as food preservatives as it has the antimicrobial activity and substitutes. Chitinase helps in the digestion of much chitin-rich foods like a mushroom¹⁰.

Sources of chitinase enzymes:

It has been found that chitinolytic enzymes are widely distributed in nature in many living organisms like bacteria, fungi, viruses, insects, some plant species, animals and in humans. In each organism, they have played different roles which help in the survival of the particular source^1,5,11,12.

Chitinase from bacteria:

Many studies have been done based on the chitinase-producing bacteria. Based on the major studies here explains some of the chitinase-producing bacteria. Streptomyces sp., Aeromonas sp., Chromobacterium sp., Klebsiella sp., Pseudomonas sp., Vibrio sp., Arthrobacter sp., Beneckea sp., Clostridium sp. and Serratia sp.¹¹. Xia et al has discovered a novel isolate of Serratia marcecens XJ-01 for the purification and optimization of chitinase. In this study, it was found that the optimum temperature for the production of the chitinase is 32 °C and pH is 8¹³. Urszula et al have purified chitinolytic enzyme from a rhizosphere strain of Stenotrophomonas maltophilia MUJ which has a strong antagonist effect towards phytopathogenic fungi such as Fusarium sp., Rhizoctonia sp. and Alternaria sp. The enzyme showed high activity at a temperature of 45 °C and pH of 6.8¹⁴.

Liang Xiao et al isolated a gram-positive bacteria Basillus strain MY75 from Shan Dong province of China which secretes a higher amount of extracellular chitinase¹⁵. Another group Hasnah Natsir et al have studied the production and characterization of chitinase from a Bacillus sp. in Sulli hot spring. The optimization study was done and it was found to be 60 °C temperature and 7.0 pH after 2 hours of incubation¹⁶. Lan et al have studied the cloning, expression, and characterization of chitinase ChiA from Aeromonas hydrophila¹⁷.

In another study by Saima et al, a novel chitinolytic bacteria Aeromonas species was isolated and found that the activity of the enzyme is higher at the optimized temperature between 22 °C to 40 °C¹⁸. Agustinus et al have done their work in the production of chitinase from the seafood waste industries. Bacillus sp. K29-14 was isolated from the crustacean waste and have found that the enzyme production was higher when compared to the colloidal chitin system^19. It has been studied that the chitinase enzyme is widely distributed in bacteria such as Serratia sp., Chromobacterium sp., Klebsiella sp., Pseudomonas sp., Clostridium sp., Vibrio sp., Arthrobacter sp., Aeromonas sp., Streptomyces sp.²⁰. Tzu-Wen Lianga,b et al described the purification and characterization of the chitinase and chitosanase, co-induced by shrimp shell powder from a strain in soil B. cereus TKU030²¹. Isurang Tantimavanich has studied on the Bacillus licheniformis TP-1 gene which produces more than one chitinases namely Chi68, Chi62, and Chi50²². An endotype chitinase chiA was cloned from Pseudoalteromonas sp. DL-6, by Xiaohui Wang which showed very high chitinase activity²³. Some of the chitinase producing bacteria are discussed in table 1.

Chitinase from seaweeds:

A Study from Nihon University, Japan showed the distribution of chitinase enzymes in seaweeds. 30 species of Rhodophyta and 1 species of Phaeophyta showed chitinase activity. Chitinases from seaweeds also play a similar role of chitinase as in higher plants³⁵.

San-Mun-Han and team³⁶ studied and identified two chitinolytic enzymes from the photosynthetic green algae Chlamydomonas renhardii for the first time. They have also studied on a novel chitinase from another photosynthetic green algae Chlorella vulgaris.

Kazuya et al.³⁷ purified chitinase isozyme (Chi A, B, and C) from red algae Chondrus verrucosus. It was found that the optimum temperature and pH for the chitinase production were at 80 °C and pH 2.0 for Chi A and C and 70 °C and pH 1 for Chi B. Chitinolytic enzymes from 98 different kinds of algae were investigated. It has been described that the isozyme Chi A, B, and C are similar to that of the yam tuber chitinases E, F, and H1 studied by Arakane in 2000³³.

Sang-Mun-Han et al described the expression and characterization of a chitinolytic enzyme in Chlorella vulgaris, green unicellular photosynthetic algae. The chitinase extracted from C. vulgaris plays a major role in the catabolism of chitin³⁶. Ali Mohammed et al have studied on the chitinase A gene from Chlorella species which is a virus sensitive strain for the first time³⁸.

Production of chitinase:

Production of chitinase is mainly influenced by the effect of temperature, pH and the growth time of the microorganism. The addition of carbon sources like glucosamine, chitobiose, etc. also induces the chitinase production. Chitinase production in the solid-state fermentation has gained much importance nowadays²⁰.

The other important methods for the microbial chitinase production are fed-batch fermentation, continuous fermentation, liquid batch fermentation etc. It has been reported that yeast has a negative effect on chitinase production. Many inhibitors like oxazolinium, allosamidin etc. have negative effects on chitinase activity⁵. The different process has been mentioned for the enrich production of chitinase-like monoculture, co-culture, recombinant cells and immobilized cells. Chitinase can be produced in a larger quantity by using recombinant microbes like recombinant E. coli.

Jing Wang and team purified two chitinases EcChi1 and EcChi2 from the hepatopancreas of the ridge tail white prawn Exopalaemon carinicauda using ion-exchange resin chromatography and gel filtration. The optimum temperature and pH of EcChi1 were 37 °C and pH 4.0, respectively³⁹. Valerie Vasseu et al have mutagenized a wild strain of the fungus Aphanocladium album by UV irradiation to get chitinase-overproducing mutants and found a 2.5-fold increase in maximal extracellular chitinase activity in this mutant⁴⁰.

Optimization of chitinase production:

For an industrial fermentation process for the production of enzymes, optimization of the media is much important as it affects the formation, concentration, and yield of a particular product⁴¹. Using different classical and statistical methods, the chitinase enzyme production is optimized.

Gasmi Meriem has optimized the chitinase production from Streptomyces griseorubens C9 by using the statistical methods of Plackett–Burman and Box–Wilson response surface methodology. Using this method, it was found that maximum chitinase was produced in media containing 2% colloidal chitin, 0.47% syrup of date, 0.25 g/L yeast extract and 1.81 g/L K₂HPO₄, KH₂PO₄ 42. Seetha Ram Kotra and team has studied the optimization of chitinase production from Streptomyces heteromorphus 4075 by using the conventional and statistical method. In this one factor at a time method was used to optimize the nutrient contents of the media and the temperature⁴³.

In a study by Noshin Tasharrofi, for the production of chitinase from Bacillus pumilus the several medium components effects were calculated by the Plackett-Burman design and they have found that chitin and yeast extract showed a positive effect on enzyme production while MgSO₄ and FeSO₄ had a negative effect. Box-Behnken response surface methodology was used to determine the optimum values⁴⁴. Crustaceans waste powder and colloidal chitin usage in different concentrations were used to optimize the chitinase production by a bacterium, Bacillus sp. K29-14 in a study by Agustinus et al found that the optimal chitinase production (4.6 U/mL) was achieved with the combined substrate of 0.5% on day 8¹⁹.

The pH 6 and temperature 30 °C were found to be optimum with colloidal chitin as a substrate and was found to be the optimized condition for the production of chitinase from Streptomyces sp. SJKP9 from the shore soil of Dhanushkodi 45. Pantoea dispersa from sea produces chitinase and the production media was optimized by using central composite design. In another study, it was found that Bacillus subtilis showed highest chitinolytic activity in Luria Bertaini Broth at pH 7.0 and temperature 35 ˚C after four days of incubation^46,47.

S.M, Akhir found that optimization of the fermentation medium for the production of chitinase from Bacillus licheniformis using Response Surface Methodology showed a 5.4-fold increase in enzyme activity than unoptimized one. Response Surface Methodology is a low-cost tool for the optimization and aimed at better enzyme production. Chitinase production by Paenibacillus sp. D1 and its optimization using statistical method showed a 2.56-fold increase in its activity^48,49,50^.

Application of chitinase:

Chitinolytic enzymes have a wide range of application in many fields. It plays a vital role in the field of medical science. One of its major and important application is chitinase is an effective biocontrol agent against pathogenic microorganisms.

Single cell protein:

Revah-Moiseev and Carrod in 1981 have mentioned the bioconversion of chitin to yeast single cell protein from shell-fish waste³. There are some fungi involved in the single cell protein production like Saccharomyces cerevisiae, Candida tropicalis, Hansenula polymorpha, and Myrothecium verrucaria¹¹. Growth yield, total protein, and nucleic acid content are the three major criteria for the production of single cell protein²⁰. In a study, it was found that the best result for the production of single cell protein was obtained when the protein content was >60% and nucleic acid was 1-3% in S. cerevisiae⁵¹. Xu Qina et al studied on the development of efficient and stable silica nanoparticles to use as a chitinase delivery system in biocontrol applications. The chitinase was isolated from B. thuringiensis⁵²^.

Biocontrol agent:

Many plants have been reported the induction of PR-protein like chitinases, proteinases etc., during the local invasion of pathogens. As the pathogenic fungus which invades the plant contains chitin in its cell wall, the chitinase present in the plant can degrade the chitin and can be a better defense. Roberts and Selitrennikoff have studied the chitinase from grains of wheat, barley and maize inhibit the hyphal elongation of the pathogenic fungi⁵¹. Chitinase can be found generally in plants as a defense mechanism against pathogen¹. Chitinase can act as a target for biopesticides and the chitinase inhibitor itself is a potent biopesticide³.

Chitin hydrolyzing enzymes are used as a biological control agent as it degrades the chitin present in the fungal cell walls and the insect’s cuticle. Chitinase isolated from the soil bacteria have antagonistic activity towards many pathogenic fungi. Subramanian et al in 2012 described the antifungal activity of chitinase from Streptomyces sp. towards Candida sp. Chitinase plays an important role in mycoparasitism also. Chitinase can be used as an additive in fungicides and insecticides to improve the activity and to reduce the concentration of chemical pesticide content ².

Urszula et al have identified and characterized a chitinase from Stenotrophomonas maltophilia, which has the antagonistic effect towards the following phytopathogenic fungi: Fusarium, Rhizoctonia and Alternaria¹⁴. H. Banani et al have described the role of chitinase from Metschnikowia fructicola in the control of brown rot disease. The results confirm that MfChi chitinase has an excellent antifungal activity to control Monilinia species, present as postharvest pathogens not only on stone fruits but also on other fruits such as apples and pears⁵³.

A positive correlation of the inhibitory activity of the chitinolytic enzyme on pathogenic fungi like R. stolonifer, B. squamosa, P. aphanidermatum, A. niger has been studied. These studies proved the use of recombinant chitinase in the protection of plants and crops from pathogenic microbes⁵⁴.

Wafaa M. Haggag purified the industrial chitinase enzyme from Streptomyces and used against different plant pathogens like Rhizoctonia solani, Fusarium oxysporum, Alternaria alternate, Aspergillus niger, Aspergillus flavus, Sclerotinia scleotiorum, Phytophthora parasitica and Botrytis cinerea. This study showed that the chitinase was optimally active at pH of 6.0 to7.0 and at 30℃. Chitinase is found to inhibit the growth of almost all phytopathogenic fungi⁵⁵.

Chitooligosaccharide production:

Chitinase is used for the production of chitooligosaccharides, which is an antibacterial agent, elicitors of lysozyme inducers and immunoenhancers³. Chitooligosaccharides can act as elicitors of plant defense, signaling for root nodule formation and is useful in medicine also²⁰. Chitohexaose and chitoheptaose show antitumor activity. GlcNAc itself is used as an anti-inflammatory drug⁵¹. Transglycosylation activity of some endochitinase is also useful for the production of chitooligomer. Purified N-acetylglucosaminidase of N. orientalis shows the transglycosylation activity.

The transglycosylation activity of chitinolytic enzymes is exploited for the synthesis of desired chitin oligomers and their derivatives. Enzymatic synthesis of chitooligosaccharides from small building blocks, and protein engineering technology for chitin-related enzymes has been considered as the most challenging part of its industrial application. Enzymatic hydrolysis of chitin using chitinase is the preferable method to chemical methods for the production of oligosaccharides ⁵⁶. An acidic chitinase purified from seeds of black soybean is an endo-splitting chitinase with short substrate cleavage activity shows the application in the production of N-acetyl Chitooligosaccharides⁵⁷.

Prognostic marker:

N. Ngernyuang et al have studied the establishment of glycoprotein 3-like 1 as a prognostic marker and therapeutic target for cervical cancer⁵⁸. Chitinase 3-Like-1 level increases up to 84% in inflammatory bowel disease. Thus, it can be used as biomarker in the diagnosis of particular disease⁵⁹.

Antifungal agent:

Massimiliano Fenice et al has been worked on the fungal chitinase in assistance with high hydrostatic pressure, which inhibits the growth of food spoiling mold Mucor plumbeus⁶⁰. Farag. M Aida et al have studied the antifungal activity of chitinase from Aspergillus terrus against pathogenic fungus. The above study explains the potential use of chitinase from A. terrus in biotechnological aspects⁶¹.

Another study was done by N.H. Zeki et al on the antifungal activity of chitinase from Serratia marcescens which has been isolated from fresh vegetable⁶². Haritha et al in another work has purified and characterized chitinase from Citrobacter freundii strain which shows antifungal activity towards pathogenic fungi A. flavus MTCC 2798 and A. niger MTCC 9652⁹.

Waste management:

Chitinous waste from the marine organisms can be converted into useful products by using recombinant chitinase^1,2. Yi Li et al have studied the algicidal activity of chitinase- producing bacteria on marine diatoms⁶³. B. Krishnaveni and R. Ragunathan have studied biodegradation of marine waste in favor of the production of highly active chitinase by Aspergillus species isolated from marine soil. This study shows a significant effect on the bioremediation of marine wastes ⁶⁴.

Medical application:

As chitinase has antifungal property it can be used as an additive in antifungal drugs. It is a naturally occurring antimicrobial agent ⁶⁵. Chitinase can be used as a biomarker and it can be used in anti-inflammatory drugs ¹. Chitinases are also used in anti-cancer therapy¹¹. Chitinase 3-like 3 (Ym1), chitinase 3-like 4 (Ym2) levels are found to be decreased after dexamethasone treatment in allergic asthma. Thus, it can be considered as a diagnostic marker in medical field 66. Having unique properties like antimicrobial, bio-flocculant or even amino-sugar (NAG) production which is considered to be most important in medical and pharmaceutical fields.

Food preservation:

The chitinase producing microorganisms appear to be the potential candidates for biological control of fungi which enhance the shelf life of various food commodities. Biological control strategies using chitinolytic enzymes are helpful in the preservation of foods concurrently from arthropods and fungal attacks. Fungi are the major problem as food spoilage organisms, human pathogen, and air contaminants. As the fungal cell walls contain almost 22% to 44% chitin, the chitinolytic enzymes can degrade the chitin cell wall of fungi and attain highly insoluble crystalline structures. Most of the fungal cell wall hydrolyses have chitinolytic activity⁶⁷. Chitinase enzyme has wide applications in the food industry by increasing shelf-life of fruits, in bioconversion process. In a study of chitinase from Streptomyces rubiginosus isolated from the rhizosphere of Gossypium sp. strain grown in colloid chitin broth showed the maximum chitinase activity⁶⁸.

Miscellaneous applications:

Fungal protoplast isolation:

As fungal protoplast is an important tool in enzyme localization study, the protoplast isolation by chitinase plays a critical role. The fungal cell wall is made up of complex structures of chitin and the chitinolytic enzyme is needed for its degradation for the protoplast isolation ^1,2.

Winemaking industry:

D. Le Bourse and team purified and characterized the chitinase enzyme in grape juice and wine. This study has a major role in finding the quantification of chitinase in grape wine. This helps the winemaking industry to improve the quality of wine⁶⁹.

Mosquito control:

Many works done on chitinase have proved that it can synergize the toxicity of the binary toxin against Culex quinquefasciatus. It is useful in the management of controlling mosquito larvae¹⁰.

CONCLUSION:

Chitinase has proved to be an emerging target for many applications. As many microorganisms like bacteria, fungi are involved in the production of chitinase, more focus should be given to the microbial chitinase production. Microbes are the more easily available natural sources for the easier production of chitinase. Recombinant techniques can be used to increase the production of chitinases for industrial purposes. Various kinetics and optimization techniques have to be used effectively for maximizing the production of chitinase. Protein engineering is used to produce chitinases with desired functions to meet the requirements of chitinases for treating various disorders. Altogether chitinase is an important factor in the development of improved agents and novel strategies for biological control.

ACKNOWLEDGMENT:

Author would like to thank VIT University for providing lab facility to carry out the work.

REFERENCES:

1. Rathore AS, Gupta RD. Chitinases from bacteria to human: properties, applications, and future perspectives. Enzyme Research 2015.

2. Nagpure A, Choudhary B, Gupta RK. Chitinases: in agriculture and human healthcare. Critical Reviews in Biotechnology 2014; 34(3):215-232.

3. Dahiya N, Tewari R, Hoondal GS. Biotechnological aspects of chitinolytic enzymes: a review. Applied Microbiology and Biotechnology 2006; 71(6):773-782.

4. Mohammed MH, Williams PA, Tverezovskaya O. Extraction of chitin from prawn shells and conversion to low molecular mass chitosan. Food Hydrocolloids 2013; 31(2):166-171.

5. Hamid R, Khan MA, Ahmad M, Ahmad MM, Abdin MZ, Musarrat J, Javed S. Chitinases: an update. Journal of Pharmacy and Bioallied Sciences 2013; 5(1):21.

6. Quecine M, Araujo W, Marcon J, Gai C, Azevedo J, Pizzirani‐Kleiner A. Chitinolytic activity of endophytic Streptomyces and potential for biocontrol. Letters in Applied Microbiology 2008; 47(6):486-491.

7. Venkatachalam A, Rajulu G, Thirunavukkarasu N, Suryanarayanan T. Endophytic fungi of marine algae and seagrasses: a novel source of chitin modifying enzymes. Mycosphere 2015; 6(3):345-355.

8. Meena S, Gothwal R, Saxena J, Nehra S, Mohan M, Ghosh P. Effect of metal ions and chemical compounds on chitinase produced by a newly isolated thermotolerant Paenibacillus sp. BISR-047 and its shelf-life. Int J Curr Microbiol App Sci 2015; 4(5):872-881.

9. Meruvu H, Donthireddy SRR. Purification and Characterization of an Antifungal Chitinase from Citrobacter freundii str. nov. haritD11. Applied Biochemistry and Biotechnology 2014; 172(1):196-205.

10. Muzzarelli RA, Boudrant J, Meyer D, Manno N, DeMarchis M, Paoletti MG. Current views on fungal chitin/chitosan, human chitinases, food preservation, glucans, pectins and inulin: A tribute to Henri Braconnot, precursor of the carbohydrate polymers science, on the chitin bicentennial. Carbohydrate Polymers 2012; 87(2):995-1012.

11. Roopavathi A, Vigneshwari R, Jayapradha R. Chitinase: Production and applications. Journal of Chemical and Pharmaceutical Research 2015; 7(5):924-931.

12. Deeba F, Shakir HA, Irfan M, Qazi JI Chitinase production in organisms: a Review.

13. Xia J-L, Xiong J, Zhang R-Y, Liu K-K, Huang B, Nie Z-Y. Production of Chitinase and its Optimization from a Novel Isolate Serratia marcescens XJ-01. Indian Journal of Microbiology 2011; 51(3):301-306.

14. Jankiewicz U, Brzezinska MS, Saks E. Identification and characterization of a chitinase of Stenotrophomonas maltophilia, a bacterium that is antagonistic towards fungal phytopathogens. Journal of Bioscience and Bioengineering 2012; 113(1):30-35.

15. Xiao L, Cai J, Lin Z-j, Han M-m, Chen Y-h. Identification and characteristics of a chitinase-produced Bacillus strain MY75. Journal of Biotechnology 2008; 136:S609-S610.

1. 16. Natsir H, Patong AR, Suhartono MT, Ahmad A. Production and characterization of chitinase enzymes from Sulili hot spring in South Sulawesi, Bacillus sp. HSA, 3-1a. Indonesian Journal of Chemistry 2010; 10(2):256-260.

16. Lan X, Zhang X, Hu J, Shimosaka M. Cloning, expression, and characterization of a chitinase from the chitinolytic bacterium Aeromonas hydrophila strain SUWA-9. Bioscience, Biotechnology, and Biochemistry 2006; 70(10):2437-2442.

17. Kuddus M, Ahmad I. Isolation of novel chitinolytic bacteria and production optimization of extracellular chitinase. Journal of Genetic Engineering and Biotechnology 2013; 11(1):39-46.

18. Uria AR, Chasanah E, Fawzya YN. Optimization of bacillus sp. K29-14 chitinase production using marine crustacean waste. Journal of Coastal Development 2013 8(3):155-162.

19. Islam R, Datta B. Diversity of chitinases and their industrial potential. Int J Appl Res 2015; 1:55-60.

20. Liang T-W, Chen Y-Y, Pan P-S, Wang S-L. Purification of chitinase/chitosanase from Bacillus cereus and discovery of an enzyme inhibitor. International Journal of Biological Macromolecules 2014; 63:8-14.

21. Tantimavanich S, Pantuwatana S, Bhumiratana A, Panbangred W. Multiple chitinase enzymes from a single gene of Bacillus licheniformis TP-1. Journal of Fermentation and Bioengineering 1998; 85(3):259-265.

22. Wang X, Zhao Y, Tan H, Chi N, Zhang Q, Du Y, Yin H. Characterisation of a chitinase from Pseudoalteromonas sp. DL-6, a marine psychrophilic bacterium. International Journal of Biological Macromolecules 2014; 70:455-462.

23. Karthik N, Akanksha K, Binod P, Pandey A. Production, purification and properties of fungal chitinases-a Review 2014.

24. Smitha S, Correya NS, Philip R. Marine fungi as a potential source of enzymes and antibiotics. International Journal of Research in Marine Sciences Universal Research Publications 2014; 3(1):5-10.

25. Barbosa N, Carvalho R, Santos E, Pietro R. Production and purification of chitinase by Metarhizium anisopliae Isolated from soil. Journal of Biotechnology 2010; 150:189-190.

26. Kawachi I, Fujieda T, Ujita M, Ishii Y, Yamagishi K, Sato H, Funaguma T, Hara A. Purification and properties of extracellular chitinases from the parasitic fungus Isaria japonica. Journal of Bioscience and Bioengineering 2001; 92(6):544-549.

27. Jenin GA, Babu MM, Murugan M, Murugan T. Isolation and Identification of Chitinase Producing Native Fungi From Saltpan of Puthalam, Kanyakumari District, Tamil Nadu, India. Journal of Applied Biology and Biotechnology 2016; Vol 4(03):001-005.

28. Jing Y, Zheng H, Yu X. Molecular cloning and sequencing of chitinase gene from Musca domestic. Journal of Biotechnology 2008; 136:S365-S366.

29. Kramer KJ, Muthukrishnan S. Insect chitinases: molecular biology and potential use as biopesticides. Insect Biochemistry and Molecular Biology 1997; 27(11):887-900.

30. Funkhouser JD, Aronson NN. Chitinase family GH18: evolutionary insights from the genomic history of a diverse protein family. BMC Evolutionary Biology 2007; 7(1):96.

31. Moder W, Bunk A, Albrecht A, Doostdar H, Niedz RP, McDonald RE, Mayer RT, Osswald WF. Characterization of acidic chitinases from culture medium of sweet orange callus tissue. Journal of Plant Physiology 1999; 154(3):296-301.

32. Arakane Y, Hoshika H, Kawashima N, Fujiya-Tsujimoto C, Sasaki Y, KOGA D. Comparison of chitinase isozymes from yam tuber—Enzymatic factor controlling the lytic activity of chitinases. Bioscience, Biotechnology, and Biochemistry 2000; 64(4):723-730.

33. Nasir N, Dharma A, Habazar T. The Chitinase Activity in Banana Seedling that Induce by Trichoderma spp as Resistance Responce to Fusarium Oxyporum f. sp. cubense. International Journal on Advanced Science, Engineering and Information Technology 2016; 6(3):356-360.

34. Sekiguchi J, Matsumiya M, Mochizuki A. Distribution of chitinolytic enzymes in seaweeds. Fisheries Science 1995; 61(5):876-881.

35. Han SM, Lee CG. Expression and Characterization of a Chitinolytic Enzyme in Chlorella vulgaris. 한국키틴키토산학회지 2008; 13(4):205-209.

36. Shirota K, Sato T, Sekiguchi J, Miyauchi K, Mochizuki A, Matsumiya M. Purification and characterization of chitinase isozymes from a red algae, Chondrus verrucosus. Bioscience, Biotechnology, and Biochemistry 2008; 72(12):3091-3099.

37. Ali AMM, Kawasaki T, Yamada T. Characterization of a chitinase gene encoded by virus-sensitive Chlorella strains and expressed during virus infection. Arab J Biotechnol 2007; 10(1):81-96.

38. Wang J, Zhang J, Song F, Gui T, Xiang J. Purification and characterization of chitinases from ridgetail white prawn Exopalaemon carinicauda. Molecules 2015; 20(2):1955-1967.

39. Vasseur V, Arigoni F, Andersen H, Defago G, Bompeix G, Seng J-M. Isolation and characterization of Aphanocladium album chitinase-overproducing mutants. Microbiology 1990; 136(12):2561-2567.

40. Panda BP, Javed S, Ali M. Fermentation process optimization. Research Journal of Microbiology 2007; 2(3):201-208.

41. Meriem G, Mahmoud K. Optimization of chitinase production by a new Streptomyces griseorubens C9 isolate using response surface methodology. Annals of Microbiology 2016; :1-9.

42. Kotra SR, Viharika V, Reddy KS, Kumari PS, Lakshminadh M, Rajesh K, Sivasai TG, Lokesh RS. Cost effective optimization of chitinase production using Streptomyces heteromorphus 4075: A progressive statistical approach. Pkhtunkhwa J Life Sci 2013; 3:100-119.

43. Tasharrofi N, Adrangi S, Fazeli M, Rastegar H, Khoshayand MR, Faramarzi MA. Optimization of chitinase production by Bacillus pumilus using Plackett-Burman design and response surface methodology. Iranian Journal of Pharmaceutical Research 2011; (4):759-768.

44. Jagadeeswari S, Selvam KP Optimization of Chitinase production by soil Streptomyces sp. SJKP9.

45. Gohel V, Jiwan D, Vyas P, Chhatpar H. Statistical optimization of chitinase production by Pantoea dispersa to enhance degradation of crustacean chitin waste. Journal of Microbiology and Biotechnology 2005; 15(1):197-201.

46. Karunya SK. Optimization and purification of chitinase produced by Bacillus subtilis and its antifungal activity against plant pathogens. International Journal of Pharmaceutical and Biological Archive 2011; 2(6).

47. Akhir S, Abd-Aziz S, Salleh M, Rahman R, Illias R, Hassan M. Medium optimisation of chitinase enzyme production from shrimp waste using Bacillus licheniformis TH-1 by response surface methods. Biotechnology 2009; 8(1):120-125.

48. Singh A, Mehta G, Chhatpar H. Optimization of medium constituents for improved chitinase production by Paenibacillus sp. D1 using statistical approach. Letters in Applied Microbiology 2009; 49(6):708-714.

49. Jholapara RJ, Mehta RS, Sawant CS. Optimization Of Cultural Conditions For Chitinase Production From Chitinolytic Bacterium Isolated From Soil Sample. Int J Pharm Bio Sci 2013; 4(2):464-471.

50. Patil RS, Ghormade V, Deshpande MV. Chitinolytic enzymes: an exploration. Enzyme and Microbial Technology 2000; 26(7):473-483.

51. Qin X, Xiang X, Sun X, Ni H, Li L. Preparation of nanoscale Bacillus thuringiensis chitinases using silica nanoparticles for nematicide delivery. International Journal of Biological Macromolecules 2016; 82:13-21.

52. Banani H, Spadaro D, Zhang D, Matic S, Garibaldi A, Gullino ML. Postharvest application of a novel chitinase cloned from Metschnikowia fructicola and overexpressed in Pichia pastoris to control brown rot of peaches. International Journal of Food Microbiology 2015; 199:54-61.

53. Yan R, Hou J, Ding D, Guan W, Wang C, Wu Z, Li M. In vitro antifungal activity and mechanism of action of chitinase against four plant pathogenic fungi. Journal of Basic Microbiology 2008; 48(4):293-301.

54. Haggag WM, Abdallh E. Purification and characterization of chitinase produced by endophytic Streptomyceshygroscopicus against some phytopathogens. Journal of Microbiology Research 2012; 2(5):145-151.

55. Jung W-J, Park R-D. Bioproduction of chitooligosaccharides: present and perspectives. Marine Drugs 2014; 12(11):5328-5356.

56. Chang Y-M, Chen L-C, Wang H-Y, Chiang C-L, Chang C-T, Chung Y-C. Characterization of an acidic chitinase from seeds of black soybean (Glycine max (L) Merr Tainan No. 3). PloS one 2014; 9(12):e113596.

57. Ngernyuang N, Francescone RA, Jearanaikoon P, Daduang J, Supoken A, Yan W, Shao R, Limpaiboon T. Chitinase 3 like 1 is associated with tumor angiogenesis in cervical cancer. The international journal of Biochemistry and Cell Biology 2014; 51:45-52.

58. Fengming Y, Jianbing W. Biomarkers of inflammatory bowel disease. Disease Markers 2014.

59. Fenice M, Di Giambattista R, Leuba J-L, Federici F. Inactivation of Mucor plumbeus by the combined actions of chitinase and high hydrostatic pressure. International Journal of Food Microbiology 1999; 52(1):109-113.

60. Aida FM, Al-Nusarie S, Taghreed S. Production, optimization, characterization and antifungal activity of chitinase produced by Aspergillus terrus. African Journal of Biotechnology 2014; 13(14).

61. Zeki N, Muslim S. Purification, characterization and antifungal activity of chitinase from Serratia marcescens isolated from fresh vegetables. Ibn Al-Haitham J Pure Appl Sci 2010; 23(1):13-25.

62. Li Y, Lei X, Zhu H, Zhang H, Guan C, Chen Z, Zheng W, Fu L, Zheng T. Chitinase producing bacteria with direct algicidal activity on marine diatoms. Scientific Reports 6 2016.

63. Krishnaveni B, Ragunathan R. Chitinase production from marine wastes by Aspergillus terreus and its application in degradation studies. International Journal of Current Microbiology and Applied Sciences 2014; 3(1):76-82.

64. Vellard M. The enzyme as drug: application of enzymes as pharmaceuticals. Current Opinion in Biotechnology 2003; 14(4):444-450.

65. Haenen S, Clynen E, Nemery B, Hoet PH, Vanoirbeek JA. Biomarker discovery in asthma and COPD: Application of proteomics techniques in human and mice. EuPA Open Proteomics 2014; 4:101-112.

66. Fuglsang CC, Johansen C, Christgau S, Adler-Nissen J. Antimicrobial enzymes: applications and future potential in the food industry. Trends in Food Science and Technology 1995; 6(12):390-396.

67. Jha S, Modi HA, Jha CK. Characterization of extracellular chitinase produced from Streptomyces rubiginosus isolated from rhizosphere of Gossypium sp. Cogent Food and Agriculture 2016; 2(1):1198225.

68. Le Bourse D, Conreux A, Villaume S, Lameiras P, Nuzillard J-M, Jeandet P. Quantification of chitinase and thaumatin-like proteins in grape juices and wines. Analytical and Bioanalytical Chemistry 2011; 401(5):1541.

69. Salam M, Dahiya N, Sharma R, Soni S, Hoondal G, Tewari R. Cloning, characterization and expression of the chitinase gene of Enterobacter sp. NRG4. Indian Journal of Microbiology 2008; 48(3):358-364.

70. Zarei M, Aminzadeh S, Zolgharnein H, Safahieh A, Daliri M, Noghabi KA, Ghoroghi A, Motallebi A. Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A. Brazilian Journal of Microbiology 2011; 42(3):1017-1029.

71. Kim K-J, Yang Y-J, Kim J-G. Purification and characterization of chitinase from Streptomyces sp. M-20. Journal of Biochemistry and Molecular Biology 2003; 36(2):185-189.

72. Annamalai N, Giji S, Arumugam M, Balasubramanian T. Purification and characterization of chitinase from Micrococcus sp. AG84 isolated from marine environment. African Journal of Microbiology Research 2010; 4(24):2822-2827.

73. Konagaya Y, Tsuchiya C, Sugita H. Purification and characterization of chitinases from Clostridium sp. E‐16 isolated from the intestinal tract of the South American sea lion (Otaria flavescens). Letters in Applied Microbiology 2006; 43(2):187-193.

74. Wang S-L, Chang W-T. Purification and characterization of two bifunctional chitinases/lysozymes extracellularly produced by Pseudomonas aeruginosa K-187 in a shrimp and crab shell powder medium. Applied and Environmental Microbiology 1997; 63(2):380-386.

75. Kang SC, Park S, Lee DG. Purification and characterization of a novel chitinase from the entomopathogenic fungus, Metarhizium anisopliae. Journal of Invertebrate Pathology 1999; 73(3):276-281.

76. Asran-Amal A, Moustafa-Mahmoud S, Sabet K, El Banna O. In vitro antagonism of cotton seedlings fungi and characterization of chitinase isozyme activities in Trichoderma harzianum. Saudi Journal of Biological Sciences 2010; 17(2):153-157.

77. Zhu Y, Pan J, Qiu J, Guan X. Isolation and characterization of a chitinase gene from entomopathogenic fungus Verticillium lecanii. Brazilian Journal of Microbiology 2008; 39(2):314-320.

78. Patil NS, Jadhav JP. Penicillium ochrochloron MTCC 517 chitinase: An effective tool in commercial enzyme cocktail for production and regeneration of protoplasts from various fungi. Saudi Journal of Biological Sciences 2015; 22(2):232-236.

79. Tweddell RJ, Jabaji-Hare SH, Charest PM. Production of chitinases and β-1, 3-glucanases by Stachybotrys elegans, a mycoparasite of Rhizoctonia solani. Applied and Environmental Microbiology 1994; 60(2):489-495.

80. Vannini A, Caruso C, Leonardi L, Rugini E, Chiarot E, Caporale C, Buonocore V. Antifungal properties of chitinases from Castanea sativa against hypovirulent and virulent strains of the chestnut blight fungus Cryphonectria parasitica. Physiological and Molecular Plant Pathology 1999; 55(1):29-35.

81. Verburg JG, Huynh QK. Purification and characterization of an antifungal chitinase from Arabidopsis thaliana. Plant Physiology 1991; 95(2):450-455.

82. Witmer X, Nonogaki H, Beers E, Bradford K, Welbaum G. Characterization of chitinase activity and gene expression in muskmelon seeds. Seed Science Research 2003; 13(02):167-178.

83. Li Y-C, Yang Y-C, Hsu JS, Wu D-J, Wu H-H, Tzen JT. Cloning and immunolocalization of an antifungal chitinase in jelly fig (Ficus awkeotsang) achenes. Phytochemistry 2005; 66(8):879-886.

Received on 28.06.2017 Modified on 20.08.2017

Research J. Pharm. and Tech 2018; 11(9): 4200-4208.

DOI: 10.5958/0974-360X.2018.00770.9

S. N	Plant	Optimized pH	Optimized Temperature (°C)	Application	Reference
1	Cabbage	6	60	-	57
2	Castanea sativa	-	-	Antifungal activity	81
3	Arabidopsis thaliana	8.7	-	Antifungal property	82
4	Muskmelon	-	30	Antifungal property	83
5	Ficus awkeotsang	-	-	-	84
6	Citrus sinensis	-	-	-	32
7	Yam tuber	-	-	-	33
8	Banana seedling	-	-	Suppress fusarium wilt disease	34

SN	Bacteria	Source	Optimized pH	Optimized Temperature (ͦ C)	Application	Reference
1	Enterobacter sp.	soil	5.5	45	Anti-fungal activity, fungal protoplast isolation, chitobiose production	70
2	Serratia marcescens	Waste water, soil	3-9	45	Antifungal activity	71
3	Streptomyces sp. M-20	-	4-8	40	Antifungal activity	72
4	Micrococcus sp. AG84	Marine environment	8	35	-	73
5	Clostridium sp. E-16	Intestinal tract of sea lion	5-7		Degradation of chitin in intestinal tract of sea lion	74
6	Pseudomonas aeruginosa	Shrimp shell	6-9	40-60	Lysozyme activity, antibacterial activity	75
7	Aeromonas hydrophila	Fresh water	-	-	ChiA production	17
8	Chromobacterium sp.	-	-	-	-	20
9	Arthrobacter sp.	-	-	-	-	20
10	Klebsiella sp.	-	-	-	-	20
11	Clostridium sp.	-	-	-	-	20
12	Vibrio sp.	-	-	-	-	20
13	Pseudoalteromonas sp.	Marine environment	-	-	High chitinase activity	23
14	Bacillus licheniformis	-	-	-	Chi68, Chi62, and Chi50 production	22
15	Beneckea sp.	-	-	-	-	20
16	Stenotrophomonas maltophilia	rhizosphere of winter wheat	-	-	-	14
17	B. cereus	soil	-	-	-	21

S.N	Fungi	Source	Optimized pH	Optimized Temperature (ͦ C)	Application	Reference
1	Metarhizium anisopliae	-	5	-	High chitinolytic activity	76
2	Trichoderma harzianum	Cotton roots	6	30	Antifungal activity	77
3	Verticillium lecanii	citrus whitefly Dialeurodes citri	7	40	Pest resistance	78
4	Penicillium ochrochloron	MTCC	-	37	fungal protoplast isolation	79
5	Stachybotrys elegans	soil	4-5	-	Antifungal activity	80
6	Trichoderma sp.	-	--		Chitinase production	24
7	Agaricus sp.	-	-	-	Chitinase production	24
8	Penicillium sp.	-	-	-	Chitinase production	24
9	Lecanicillium sp.	-	-	-	Chitinase production	24
10	Neurospora sp.	-	-	-	Chitinase production	24
11	Mucor	-	-	-	Chitinase production	24
12	Beauveria sp.	-	-	-	Chitinase production	24
13	Lycoperdon sp.	-	-	-	Chitinase production	24
14	Aspergillus sp.	-	-	-	Chitinase production	24
15	Myrothecum sp.	-	-	-	Chitinase production	24
16	Conidiobolus sp.	-	-	-	Chitinase production	24
17	Metharhizum sp.	-	-	-	Chitinase production	24
18	Stachybotrys sp.	-	-	-	Chitinase production	24