INTRODUCTION:

Glucans are ring-shaped diverse glucose molecules interlinked by glycosidic bonds. The glycosidic bond classifies Glucans into two categories, alpha (α-) are polysaccharides formed by D-glucose molecules joined by alpha-glycosidic linkages, and beta (β-) β-D-glucose polysaccharides interlinked by beta (β-) glycosidic linkages as shown in Fig.1. β-Glucans are high molecular weight substances usually isolated from the cell walls of many prokaryotic and eukaryotic organisms¹. All β- Glucans glycosidic linkages join D-glucose polysaccharide chains, but all β-D-glucose linkages don't fulfill the requirement desirable for β-Glucans formation². The most widespread β-Glucans backbone linkages such as β- D-(1→3), β- D-(1→4), and β- D-(1→6) are reported in many prokaryotes and eukaryotes³. β-Glucans have more nutraceutical significance than α-Glucansdue to which their market size has reached US$ 501 Million in the year 2023 and is predicted to reach US$ 734 Million by the year 2028, showing a compound annual growth rate (CAGR) of 7.64% between 2023 and 2028⁴. The review explores β-Glucans diverse sources, synthesis pathways for prokaryotes and eukaryotes, chemical and physical extraction methods. Later, the significance of various techniques in identifying the structure of β-Glucans is also discussed.

Fig. 1. Types and subtypes of Glucans i.e. Alpha (α-) and Beta (β-Glucans)

Sources of β-Glucans:

The β-Glucans exist in both prokaryotes and eukaryotes. The percentage of β-Glucans in prokaryotic and eukaryotic organisms is illustrated in Fig. 2. Some significant sources of β-Glucans are described below such as:

Prokaryotes:

β-Glucans are reported in numerous positive and gram-negative organisms such as Lactobacillus spp, Agrobacterium spp, Bacillus subtilis,andXanthomonas campertis. Lactobacillus fermentum (Lb. fermentum) is a Gram-positive bacterium that exists in soil, plants, and animals and is responsible for boosting immunity and guard against gastrointestinal and upper respiratory illnesses. It is used as an antibiotic substitute and food preservative⁵. Agrobacteria spp is a class of soil bacteria used as a model organism in genetic engineering^6, Bacillus subtilis is present in the human gut and fermented foods and is defined as a potential probiotic organism used to treat intestinal disorders⁷. Xanthomonas campestris is a Gram-negative bacteria, present in soil utilized for the industrial production of xanthan gum⁸. Hence extraction of β-Glucans from non-pathogenic organisms highlights the applicability of these organisms along with their metabolite.

Eukaryotes:

β-Glucans are present in the highest concentration in the eukaryotic organism Euglena⁹, a single-cell eukaryotic organism possessing strong antibiotic and expectorant properties. In cereals, β-Glucans the percentage is comparatively lower but using them as a β-Glucans source is reported useful for regulating the concentration of cholesterol and glucose in blood¹⁰.

Fig. 2. Sources of β-Glucans and their concentration in Prokaryotes and Eukaryotes.

Biosynthesis of β-Glucans:

Various β-Glucans biosynthesis mechanisms are reported in prokaryotes and eukaryotes, such as:

Prokaryotes:

In prokaryotes, the biosynthesis of β-Glucans involves a cascade of enzymatic reactions. Various mono and disaccharides i.e., glucose and maltose have been proposed as potential starting points. These reactions are observed mainly in the inner surface of the cytoplasmic membranes. Different types of transporters play a crucial role. The steps involved in the biosynthesis of β-Glucans in prokaryotic organisms e.g., Lactobacillus brevis¹¹ are as shown in Fig. 3:

1. Transport of maltose:

Maltose from the extracellular membrane enters the intracellular membrane with the help of MFS-Transporter.

2. Formation of Glucose -1-phosphate:

Inside the cell, maltose is transformed into Glucose -1-phosphate with the enzyme maltose phosphorylase.

3. Formation of Glucose -6-Phosphate:

Glucose -1-phosphate is converted to Glucose -6-phosphate in the presence of enzyme β-phosphoglucomutase.

4. Synthesis of UDP-glucose:

Glucose-1-phosphate in the presence of enzyme UTP-Glucose-1-phosphate uridyl transferase is responsible for the formation of UDP-Glucose. In the reaction, UTP is converted to UDP.

5. Formation of β-Glucan precursors:

UDP-glucose is converted to β-Glucan precursors, such as laminaribiose and sophorose by the action of glycosyltransferase-2. In the reaction also, UTP is converted to UDP.

6. Polymerization of β-Glucans:

β-Glucan synthase enzymes are required to form long chains of β-Glucans to polymerize the β-Glucan precursors. These chains are extruded through the cytoplasmic membrane and merged into the cell wall.

7. Modification of β-Glucans:

After polymerization, the β-Glucans may undergo further modifications, such as branching or cross-linking, to increase their strength and stability.

Fig. 3. Metabolic pathway of the β-Glucans biosynthesis in Prokaryotes (e.g. lactobacillus brevis) from genomic data and proteomic analysis. 1. MFS-Transporter is a major facilitator superfamily which helps Maltose from the extracellular membrane enters the intracellular membrane. 2. Maltose phosphorylase, Glucokinase, UTP-Glucose-1-phosphate uridyltransferase, Nucleoside-diphosphate kinase, Gtf-2 i.e.Glycosyltransferase -2 are enzymes responsible for the synthesis of β-Glucans.

Eukaryotes:

In eukaryotes, such as fungi and plants, the biosynthesis of β-Glucans is intricate and requires enzymes. Monosaccharide i.e., glucose served as a potential starting point. The biosynthetic pathway (e.g., Saccharomyces cerevisiae¹² is broadly divided into the following steps and summarised in Fig. 4.

1. Transport and Phosphorylation of Glucose:

Glucose from the extracellular membrane enters the intracellular membrane with the help of a transporter. Later it is transformed into Glucose-6-phosphate by the enzyme hexokinase (HXK1). It is then transformed to glucose-1-phosphate by phosphoglucomutase (Pgm2).

2. Synthesis of nucleotide sugar precursors:

The next footstep in the biosynthesis of β-Glucans in eukaryotes is the formation of nucleotide sugar precursors, such as UDP-glucose from UTP and glucose-1-phosphate, formed by a series of enzymatic reactions.

3. Assembly of β-glucan polymers:

In this step, nucleotide sugar precursors are used as substrates by specific β-Glucan synthase enzymes. It is an important step required to assemble β-Glucan polymers. The β-Glucan synthase enzymes promote the transfer of glucose or other sugar units from the nucleotide sugar precursors to the growing β -Glucan chains¹².

4. Modification and transport of β-Glucans:

The β-Glucans polymers are modified by enzymes and transported to the cell wall of the organism. Enzymes add side chains to the β-Glucans chains. Once modified, the β-Glucans polymers are transported to the cell wall and incorporated into the matrix. The polymerization of β-Glucans from UDP-glucose takes place by the glucan synthases (Fks1, Fks2, and Fks3) in cooperation with Rho1. As the β-Glucans biosynthetic pathway in eukaryotes is regulated by a multifaceted network of enzymes and signalling pathways, mutations in these pathways can lead to defects in β-Glucans biosynthesis and result in developmental abnormalities that can lead to diseases¹².

Fig. 4. Metabolic pathway of the β-Glucan biosynthesis in Eukaryotes (e.g. Saccharomyces cerevisiae). 1. Fks1, Fks2, and Fks3 are glucan synthases that produce β- D-(1→3) glucan from UDP-glucose. 2. Gtf and Crd are exogenous glucan synthases that produce β- D-(1→3) glucan from UDP-glucose. 3. From UDP glucose, GsmA produces β- D-(1→6) Glucan.

Non-biological Synthesis of β–Glucans:

β-Glucans can also be synthesized using non-biological techniques in which hydrolysis of substrate or product can be avoided easily with improved size, properties, and stable β-Glucans. The non-biological methods involved in the synthesis of the β-Glucans are as follows

1. Enzymatic synthesis:

Enzymatic synthesis is one of the most widely used methods to form β-Glucans. Some of the enzymes that actively participate in the biosynthesis of β-Glucans are such as:

Phosphoglucomutase:

Phosphoglucomutase is involved in the transfer of a phosphate group on the α-D-glucose molecule from 1 to 6 position in the accelerative direction or the 6 to 1 position in the opposite direction¹².

Glycosyltransferases:

Glycosyltransferases (GTs) are involved in the synthesis of glycosidic bonds between an activated sugar donor and a non-activated acceptor carbohydrate. This activity can occur on biomolecules such as monosaccharides, disaccharides, oligosaccharides, lipopolysaccharides, and peptidoglycan¹³.

Glycosynthases:

Glycosynthases are the first non-natural, biosynthetic enzyme capable of in vitro production of β-Glucans¹³. Numerous glycosynthases participate in synthesizing β-Glucans. For example, β-1, 3 glycosynthases effectively utilized to synthesize β-D-(1→3) Glucans β-1, 4 glycosynthases also known as Glycoside phosphorylases catalyze the splitting of glycosidic bonds liberating sugar 1-phosphate and shorter glycans in the presence of inorganic phosphate¹³. β-1, 2 glycosynthases improve the availability of certain phenolic compounds by glycosylation¹⁴.

UDP-glucose pyrophosphorylase:

UDP-glucose pyrophosphorylase is responsible for the interconversion between Glucose-1-phosphate (Glu-1-P) and Uridine triphosphate (UTP) to UDP-glucose (UDP-Glu) and inorganic pyrophosphate (PP_i)¹⁵.

Glucan synthases:

Glucan synthases synthesize β-D-(1→3) Glucan from UDP-glucose. β-D-(1→3) Glucan synthase is a glucosyltransferase enzyme participating in the formation of β-Glucans in fungi¹⁶.

2. Chemical synthesis:

Chemical synthesis is another approach to the non-biological synthesis of β-Glucans, which involves methods, such as acid hydrolysis or oxidation of cellulose. In acid hydrolysis, cellulose is treated with acid to break down the cellulose fibers into shorter β-Glucans chains. However, in oxidation, the cellulose fibers are treated with oxidizing agents to create carboxyl groups that play an essential role in linking and forming chains of β-Glucans¹⁷.

3. Physical synthesis:

Physical methods such as ultrasonication or high-pressure homogenization have also been used in synthesizing β-Glucans. These methods can break down larger polysaccharides into smaller β-Glucans chains, which can then be further processed or modified. This is an effective pre-treatment before autolysis of Saccharomyces cerevisiae cells for improved production of yeast extract¹⁸.

Extraction of β-Glucans:

Various extraction techniques play a substantial role in extracting β-Glucans as discussed in detail in Table 1.

1. Enzymatic Extraction:

Enzymatic extraction can be performed by using various enzymes to extract β-Glucans from different sources. Enzymatic extraction with α-amylase is utilized for the isolation of β-Glucans from grains like barley and oats¹⁹. Whereas Enzymatic extraction with β-glucanase, Lipase, and protease is utilized for Yeast Saccharomyces cerevisiae²⁰.

2. Chemical Extraction:

Chemical Extraction can be performed as Acidic extraction, alkaline extraction, and Combined Acidic and Alkaline extraction.

Acidic extraction:

Acidic extraction with citric acid is utilized for oats and barley. Acidic extraction with perchloric acid is utilized for malt and barley grains whereas Acidic extraction with KOH is utilized for the isolation of β-Glucans from Usnea Lichen^19,20.

Alkaline extraction:

Alkaline extraction with Na₂CO₃ is utilized for extracting β-Glucans from oats and oat bran. In contrast, alkaline extraction with NaOH is utilized for the isolation of β-Glucans from various grains like barley bran, oat bran, wheat bran, and mushroom-like Macrocybe titans²⁰.

Combined Acidic and Alkaline extraction:

Combined acidic and alkaline extraction is utilized for the isolating of β-Glucans from Yeast Saccharomyces cerevisiae, red algae, and rice bran²¹.

3. Physical Extraction:

Physical extraction can be performed by Milling and sieving as well as Ultrafine grinding for oats. High-pressure homogenization, sonication, and ultrasonication for Yeast Saccharomyces cerevisiae. A pilot-scale procedure combining reverse osmosis (nanofiltration), and crossflow microfiltration is carried out for Shiitake mushroom (Lentinula edodes)²¹.

4. Water Extraction:

The water extraction technique is useful for isolating β-Glucans from barley and oats. In this technique, extraction temperature is important in the yield and purity of β-Glucans. Water extraction at a mild Temperature (< 50 ⁰C) is utilized for the isolation of barley and Oats. Water extraction at (135.0 ⁰C) is also utilized for the isolation of barley whereas by using Pressurized hot water extraction at (157.5⁰C) waxy barley can be isolated. Water extraction minimizes the usage of chemicals but it is a highly time-consuming (approximately 7 d) process^19,20,21.

Table 1. Extraction of β-Glucans

Sr. No.	Extraction methods of β-Glucans	Source of β-Glucans	Extraction Yield of β-Glucans (%)	Purity of β-Glucans (%)	References
Enzymatic Extraction
i)	Enzymatic extraction with α-amylase	Barley	5.4	83.1	19
i)	Enzymatic extraction with α-amylase	Oat	13.9	-	19
ii)	Enzymatic extraction with β glucanase, lipase, and protease	Yeast Saccharomyces cerevisiae	10.01	65	20
Chemical Extraction
a.	Acidic Extraction
i)	Acidic extraction with citric acid	Oat	6.97	-	19,20
i)	Acidic extraction with citric acid	Barley	4.65	80.4
ii)	Acidic extraction with perchloric acid	Malt and Barley grains	4.3 to 6.0	71.2 to 73.3
iii)	Acidic extraction with potassium hydroxide (KOH)	Usnea lichen	4.5 to 6.5	68
b.	Alkaline Extraction
i)	Alkaline extraction with sodium carbonate	Oat bran	8.57	48.5 to 65.99	19
ii)	Alkaline extraction with sodium hydroxide	Wheat bran	2.15 to	91.58	20
ii)	Alkaline extraction with sodium hydroxide	Barley bran, Macrocybe titans	2.51 5.6 to 11.9	73 to 77	20
c.	Combined Extraction
i)	Sodium hydroxide / Hydrogen chloride extraction	Yeast Saccharomyces cerevisiae, red algae, and rice bran	66.67	- -	21
ii)	Sodium hydroxide / Acetic acid extraction		68.18
iii)	Sodium hydroxide / Sodium hypochlorite extraction		61.82
iv)	Sodium hydroxide and Sodium hypochlorite / Dimethyl sulfoxide extraction		58.33
Physical Extraction
i)	Milling and sieving	Oat	6.72	56.2,64.3	21
ii)	Ultrafine grinding	Oat	-	56.2	21
iii)	High-pressure homogenization	Yeast Saccharomyces cerevisiae, Lentinula edodes	-	-	21
iv)	Sonication, ultrasonication	Yeast Saccharomyces cerevisiae, Lentinula edodes	-	-	21
Water Extraction
i)	Water extraction (Mild Temperature < 50 °C)	Barley	2.5 to 5.4	85 to 91.3 90.4 to 93.7	19,20,21
i)	Water extraction (Mild Temperature < 50 °C)	Oats	2.1 to 3.9	85 to 91.3 90.4 to 93.7
ii)	Water extraction (Hot 135.0 °C)	Barley	5.4	83.8
iii)	Hot water extraction (Pressurized 157.5^oC)	Waxy barley	-	50.8 to 54

In addition to conventional methods, other techniques such as microwave extraction, supercritical fluid extraction, and soxhlet-assisted extraction can also be utilized to decrease the extraction time, shorten solvent consumption, improve the yield, and boost the quality of the extract^22,23.

Characterization of β-Glucans:

The identification and characterization of β-Glucans can be performed using the following techniques as well-defined in Table 2.

1. Fourier Transform Infrared Spectrophotometry (FTIR):

Fourier Transform Infrared Spectroscopy (FTIR) spectra were performed at a resolution of 2 cm⁻¹ between 4000 cm⁻¹ and 400 cm^{−1 24}. The peaks for the –HO stretch were spotted at 3173 cm⁻¹, and 3390 cm⁻¹. For the C-C-C stretch, peaks were detected at 1608.52 cm⁻¹ and 1668.31 cm⁻¹; for the R-O-R stretch, peaks were reported at 1075.24 cm⁻¹, and 1228.57 cm⁻¹ ²⁵. Furthermore, C–O stretch peaks were observed at 2919 cm^-1 and for C–H stretch at 1076 ²⁶. Interestingly a peak at 890 cm⁻¹ highlights β-linked β-Glucans polymer^26,27

2. Ultraviolet-Visible (UV-Vis) spectrophotometry:

Ultraviolet-visible (UV-Vis) spectroscopy is used to evaluate unique UV or visible light wavelengths that β-Glucans absorb or transmit, providing information about the concentration of β-Glucan. The UV/Vis absorbance peak in natural polymers at 260 nm, highlights the presence of carbonyl groups signifying the prevalence of β-Glucans²⁸. The broad UV/Vis absorption spectrum of β-Glucans ranges from 260 to 300 nm^29,30.

3. Nuclear Magnetic Resonance (NMR) spectrophotometry:

Nuclear magnetic resonance (NMR) spectroscopy in the 3 kHz–300 GHz34 radiofrequency range is useful to detect nuclei such as 1H and 13C to evaluate structural linkages between glucose molecules in β-Glucans. Two-dimensional NMR spectroscopy is utilized for detailed structural characterization of aminated Zymosan (ZM) extracted from Saccharomyces cerevisiae³¹. It is observed that resonance between 60 ppm and 105 ppm in the 13C NMR spectrum of curdlan is attributable to the β- D-(1→3) and β- D-(1→6) linkages in the β-Glucans³². However, NMR spectra resonance peaks in barley at 104 ppm, 69 ppm, and 62 ppm confirm β- D-(1→3) and β- D-(1→6) linkages in the β-Glucans^33,34.

4. High-Performance Anion Exchange Chromatography (HPAEC):

High-performance anion Exchange Chromatography (HPAEC) investigates the structural properties of β-Glucans such as anomericity, composition, size, and linkage isomerism³⁵. The linkage ratio of soluble and insoluble triose to tetrose fraction of β-Glucans suggests differences in its composition or branching structure. Linkage ratios of barley β- Glucans were found to be in the range of 2.27-2.31, whereas in oat β- Glucans in the range of 2.38-2.39 representing β- D-(1→3), β- D-(1→4), and β- D-(1→6) glycosidic linkages in β-Glucans³⁶.

5. High-Performance Liquid Chromatography (HPLC):

High-Performance Liquid Chromatography (HPLC) detects the retention time of β-Glucans which is quantitated according to the peak area of the β-Glucans³⁷. β-Glucans of oats and edible mushroomswere found to be in the range of 2.94 - 3.16 min (377.215 mAU-487.633 mAU)³⁸. However, the retention time of β-Glucans from Lactobacillus fermentum was 9.024 min^39,40.

6. Differential scanning calorimetry (DSC):

Differential Scanning Calorimetry (DSC) thermoanalytically measures temperature-dependent differences between β-Glucans which is helpful to detect the structural stability and behavior of β-Glucans with changing temperatures. The thermal transition temperature (melting peak) of β-Glucans was observed in the range of 122⁰C - 125⁰C²⁴. The melting peak in the Eukaryotic organism Yeast was observed at 118⁰C specifying the presence of β-Glucans^41,42.

7. X-ray diffraction (XRD):

X-ray diffraction (XRD) was executed at 40 kV and 40 mA to determine physical properties such as flexibility, solubility, swelling, and tensile strength by analyzing the crystal structure of β-Glucans. Yeast β-Glucans had significantly higher crystallinity between 20% - 28.37%^43.The samples were investigated in an angular (2θ) range from 5⁰ - 65⁰ at a scanning speed of 6⁰/min. Natural polymers had an absorption at 19.6⁰whereas, Yeast β-Glucans had the strongest absorption at 20⁰, due to their polymeric structure^43,44.

Table 2. Identification and Characterization of β-Glucans

1. Fourier transform infrared spectrophotometry (FTIR) spectra of β-Glucans
Sr. No.	Sources of β-Glucans	Extraction	O-H stretch (cm⁻¹) (3000-3700)	C-H stretch (cm⁻¹)	C-C stretch (cm⁻¹) (1680–1600) and C-O stretch	(C₁–H) deformation mode, i.e., β-glycosidic bonds	(C=O) amide stretch	R-O-R stretch (cm⁻¹) (1050–1260)			References
1.	Yeast	Acid-base extraction	3412	2921	900–1200	978.24	1581	-			24,25
2.	Yeast	Water extraction	3445	2851	900–1200	978.24	1661	-			25
3.	Yeast	Alkali-acid method	3173	-	1608.52 - 1668.31 (C-C-C stretch)	885	-	1075.24 - 1228.57			25
4.	Yeast	Alkali-acid method	3390	2919 (C–H stretch)	1076 (C-O stretch)	890	-				26
2. Ultra violet-visible (UV-Vis) absorbance of β-Glucans
Sr. No.	Sources of β-Glucans			UV/Vis absorbance (nm) (260 - 300)					References
1.	Natural Natural polymers			260					28,29,30
3. Nuclear magnetic resonance (NMR) of β-Glucans
Sr.No.	Sources of β-Glucans		Linkages	Resonance spectrum (ppm) (60 -105)					References
1.	Barley		β- D-(1→3)	104					31,32,33,
2	Saccharomyces cerevisiae		β- D-(1→3)	101
3.	Barley		β- D-(1→6)	62,69
4. High-Performance Anion Exchange Chromatography (HPAEC) of β-Glucans
Sr.No.	Sources of β-Glucans		Linkages	Linkage ratios					References
1.	Barley		β- D-(1→3), β- D-(1→4), β- D-(1→6)	2.27-2.31					35,36
2.	Oats			2.38-2.39
5. High-Performance Liquid Chromatography (HPLC) of β-Glucans
Sr. No.	Sources of β-Glucans			Retention time (min)		milli-Absorbance Units (mAU)			References
1.	Oats			2.94		487.633			37,38
2.	Edible mushroom			3.16		487.633
3.	Lactobacillus fermentum			9.024		-			39,40
6. Differential scanning calorimetry (DSC) of β-Glucans
Sr. No.	Sources of β-Glucans			Melting peak (°C)					References
1.	Yeast (After decomposition)			122 – 125					24,41
2.	Yeast			118					42
7. X-ray diffraction (XRD) absorbance of β-Glucans
Sr. No.	Sources of β-Glucans			Absorption (2θ)		Crystallinity (%)	Content (%)			References
1.	Natural polymers			19.6⁰		-	-			43
2	Yeast			20⁰		20 - 28.37	77.34 - 80.14			44

In addition to traditional methods, crucial techniques like elemental analysis by inductive coupled Plasma (ICP), energy dispersive X-ray analysis (EDAX), physical characterization, and reversed-phase liquid chromatography (RF-HPLC) can also play a significant for the identification and characterization of β-Glucans^45,46.

CONCLUSION:

Glucans are diverse glucose molecules linked together by glycosidic bonds that exist in both prokaryotes and eukaryotes. In Glucans, glucose molecules may differ in the position of glycosidic bonds which classifies Glucans into two categories, alpha (α-) and beta (β-). β-Glucans have more nutraceutical significance than α-Glucans due to their regio-/stereo-selectivity. Understanding the biological and non-biological synthetic pathways of β-Glucans is important for developing strategies to modify β-Glucans synthesis. Non-biological synthesis, such as the chemical synthesis of β-Glucans, is challenging because it requires building blocks as reactants and has poor regio-/stereo-selective reactions. β-Glucans synthetic pathways could be better understood and explored by combining omics approaches (such as genomics, metabolomics, proteomics, and transcriptomics) with artificial intelligence. To date, the exact mechanism for β-Glucans synthesis is less explored. Therefore, developments in synthesis methodologies, novel catalysts, and innovative strategies will help to address these challenges and to develop better ways to synthesize β-Glucans. Various extraction techniques are used to separate β-Glucans from different sources. Water extraction is an eco-friendly technique that minimizes the usage of hazardous chemicals, but it is a highly time-consuming process. The need for optimization of non-hazardous, non-time-consuming extraction techniques, is a critical area for further research. There are multiple characterization techniques for β-Glucans identification and characterization. The challenges for biosynthesizing, extracting, and characterizing β-Glucans require interdisciplinary collaborations, standardized methodologies, and innovative approaches to bridge the knowledge gaps so that this multifunctional molecule can be used at a wider level to impart more health-beneficial impacts.

ACKNOWLEDGMENT:

The authors would like to express their heartfelt gratitude to the Department of Biosciences and Technology, Department of Pharmaceutical Sciences and Technology, Dr. Vishwanath Karad MIT World Peace University Pune, for providing all the necessary facilities to carry out the literature survey.

CONFLICTS OF INTEREST:

The authors declared no conflict of interest in the manuscript.

REFERENCES:

1. Mirończuk-Chodakowska I, Kujawowicz K, Witkowska AM. Beta-glucans from fungi: biological and health-promoting potential in the COVID-19 pandemic era. Nutrients. 2021; Nov 6; 13(11): 3960. https://doi.org/10.3390/nu13113960

2. Du B, Meenu M, Liu H, Xu B. A concise review of the molecular structure and function relationship of β-glucan. International Journal of Molecular Sciences. 2019; Aug 18; 20(16): 4032.https://doi.org/10.3390/ijms20164032

3. Stone BA. Chemistry of β-glucans. In Chemistry, Biochemistry, and Biology of 1-3 beta glucans and Related Polysaccharides 2009 Jan 1 (pp. 5-46). Academic press. https://doi.org/10.1016/B978-0-12-373971-1.00002-9

4. Market Reports (2023). Beta-glucan market report. Retrieved from Market Reports (2023). Beta-glucan market report. https://www.marketsandmarkets.com/Market-Reports/beta-glucan-market-5191796. Accessed December 20, 2023.

5. Naghmouchi K, Belguesmia Y, Bendali F, Spano G, Seal BS, Drider D. Lactobacillus fermentum: A bacterial species with potential for food preservation and biomedical applications. Critical reviews in Food Science and Nutrition. 2020; Nov 12; 60(20): 3387-99. https://doi.org/10.1080/10408398.2019.1688250 2020.

6. Zhou Y, Luo Y, Yu B, Zheng P, Yu J, Huang Z, Mao X, Luo J, Yan H, He J. Agrobacterium sp. ZX09 β-Glucan Attenuates Enterotoxigenic Escherichia coli-Induced Disruption of Intestinal Epithelium in Weaned Pigs. International Journal of Molecular Sciences. 2022; Sep 7; 23(18): 10290. https://doi.org/10.3390/ijms231810290

7. Arena MP, Spano G, Fiocco D. β-Glucans and Probiotics. American Journal of Immunology. 2017; 13(1): 34-44. https://doi.org/10.3844/ajisp.2017.34.44

8. Utama GL, Dio C, Lembong E, Cahyana Y, Balia RL. Microorganism-based β-glucan production and their potential as antioxidants. Systematic Reviews in Pharmacy 2020; Oct 1; 11: 868-73. http://dx.doi.org/10.31838/srp.2020.10.130

9. Schulze C, Wetzel M, Reinhardt J, Schmidt M, Felten L, Mundt S. Screening of microalgae for primary metabolites including β-glucans and the influence of nitrate starvation and irradiance on β-glucan production. Journal of Applied Phycology. 2016; Oct; 28: 2719-25. https://doi.org/10.1007/s10811-016-0812-9

10. Lante, A.; Canazza, E. Insight on Extraction and Preservation of Biological Activity of Cereal β-D-Glucans. Appl. Sci. 2023; 13: 11080. https://doi.org/10.3390/ app131911080

11. Bockwoldt JA, Meng C, Ludwig C, Kupetz M, Ehrmann MA. Proteomic analysis reveals enzymes for β-D-glucan formation and degradation in Levilactobacillus brevis TMW 1.2112. International Journal of Molecular Sciences. 2022; Mar 21; 23(6): 3393.https://doi.org/10.3390/ijms23063393

12. Zhou X, He J, Wang L, Wang Y, Du G, Kang Z. Metabolic engineering of Saccharomyces cerevisiae to improve glucan biosynthesis. Journal of Microbiology and Biotechnology May 2019; 29(5):758-764https://doi.org/10.4014/jmb.1812.12049

13. Bulmer GS, De Andrade P, Field RA, van Munster JM. Recent advances in enzymatic synthesis of β-glucan and cellulose. Carbohydrate Research. 2021; Oct 1; 508: 108411. https://doi.org/10.1016/j.carres.2021.108411

14. Méndez-Líter JA, Nieto-Domínguez M, Fernandez de Toro B, González Santana A, Prieto A, Asensio JL, Cañada FJ, de Eugenio LI, Martínez MJ. A glucotolerant β-glucosidase from the fungus Talaromyces amestolkiae and its conversion into a glycosynthase for glycosylation of phenolic compounds. Microbial Cell Factories. 2020; Dec; 19:1-3. https://doi.org/10.1186/s12934-020-01386-1

15. Kleczkowski LA, Decker D. Effects of magnesium, pyrophosphate, and phosphonates on the pyrophosphorolytic reaction of UDP-glucose pyrophosphorylase. Plants. 2022; Jun 20; 11(12): 1611. https://doi.org/10.3390/plants11121611

16. Chhetri A, Loksztejn A, Nguyen H, Pianalto KM, Kim MJ, Hong J, Alspaugh JA, Yokoyama K. Length specificity and polymerization mechanism of (1, 3)-β-d-glucan synthases in fungal cell wall biosynthesis. Biochemistry. 2020; Jan 3; 59(5): 682-93. https://doi.org/10.1021/acs.biochem.9b00896.

17. Ferrières V, Legentil L, Sylla B, Descroix K, Nugier-Chauvin C, Daniellou R. Chemical Synthesis of Oligo-(1 3)- -D-Glucans. Biology and Chemistry of Beta Glucan. 2013 May 24; 2:83.

18. Dimopoulos G, Tsantes M, Taoukis P. Effect of high-pressure homogenization on the production of yeast extract via autolysis and beta-glucan recovery. Innovative Food Science and Emerging Technologies. 2020 Jun 1; 62:102340.https://doi.org/10.1016/j.ifset.2020.102340

19. Maheshwari G, Sowrirajan S, Joseph B. Extraction and isolation of β‐glucan from grain sources—A review. Journal of Food Science. 2017; Jul; 82(7): 1535-45. https://doi.org/10.1111/1750-3841.13765.

20. Varelas V, Liouni M, Calokerinos AC, Nerantzis ET. An evaluation study of different methods for the production of β‐D‐glucan from yeast biomass. Drug Testing and Analysis. 2016 Jan;8(1):46-55. https://doi.org/10.1002/dta.1833

21. Pengkumsri N, Sivamaruthi BS, Sirilun S, Peerajan S, Kesika P, Chaiyasut K, Chaiyasut C. Extraction of β-glucan from Saccharomyces cerevisiae: Comparison of different extraction methods and in vivo assessment of immunomodulatory effect in mice. Food Science and Technology. 2016 Jul 28; 37: 124-30. https://doi.org/10.1590/1678-457X.10716.

22. Upadhye M, Gandhi P, Phanse M. Pharmacognostical, Phytochemical Studies and Comparative Extraction Techniques of Daucus carota. Research Journal of Pharmacognosy and Phytochemistry. 2014; 6(3): 115-7.

23. Pawar AR, Vikhe DN, Jadhav RS. Recent Advances in Extraction Techniques of Herbals-A Review. https://doi.org/10.5958/2231-5659.2020.00050.8

24. Bacha U, Nasir M, Iqbal S, Anjum AA. Nutraceutical, anti-inflammatory, and immune modulatory effects of β-glucan isolated from yeast. BioMed Research International. 2017; Aug 23; 2017.https://doi.org/10.1155/2017/8972678.

25. Indriati Ramadhani DL, Yuliani Y, Amir M. Extraction, characterization, and biological toxicity of β-glucans from Saccharomyces cerevisiae isolated from ragi. Journal of Microbial Systematics and Biotechnology. 2020; 2(2): 35-43. http://dx.doi.org/10.37604/jmsb.v2i2.62.

26. Jean A. Boutros; Andrew S. Magee; Donald Cox; (2022). Comparison of structural differences between yeast β-glucan sourced from different strains of Saccharomyces cerevisiae and processed using proprietary manufacturing processes. Food Chemistry. https://doi.org/10.1016/j.foodchem.2021.130708.

27. Kanchana S, Arumugam M. Alternative exploration of hyaluronic acid from the marine superstore. Asian Journal of Pharmaceutical Research. 2014; 4(4): 169-73.

28. Cho JH, Kim T, Yun HY, Kim HH. Facile depolymerization process of β-glucan through the use of a high-pressure homogenizer. American Journal of Research Communication. 2014; 2: 168-78.

29. Lee K, Choi Y, Kim K, Koo HJ, Choi J. Quantification of unknown nanoscale biomolecules using the average-weight-difference method. Applied Sciences. 2019; Jan 2; 9(1): 130.https://doi.org/10.3390/app9010130.

30. Yogeshwari C, Kumudha P. Spectroscopic Analysis of Bioactive Components of Ichnocarpus frutescens R. Br. Research Journal of Pharmacognosy and Phytochemistry. 2018; 10(2): 171-4. https://doi.org/10.5958/0975-4385.2018.00026.2

31. Venkatachalam G, Arumugam S, Doble M. Synthesis, characterization, and biological activity of aminated zymosan. ACS omega. 2020 Jun 23; 5(26): 15973-82. https://doi.org/10.1021/acsomega.0c01243.

32. Rong Y, Xu N, Xie B, Hao J, Yi L, Cheng R, Li D, Linhardt RJ, Zhang Z. Sequencing analysis of β-glucan from highland barley with high-performance anion exchange chromatography coupled to quadrupole time–Of–Flight mass spectrometry. Food hydrocolloids. 2017; Dec 1; 73: 235-42. http://dx.doi.org/10.1016/j.foodhyd.2017.07.006.

33. Bikmurzin R, Bandzeviciute R, Maršalka A, Maneikis A, Kalėdienė L. FT-IR method limitations for β-glucan analysis. Molecules. 2022; Jul 20; 27(14): 4616. https://doi.org/10.3390/molecules27144616

34. Sahoo MR, Varrier RR, Anithakumari R, Palanichamy G, Sundari BT, Guru B. Analytical Profiling of Saffron (Crocus sativus) using 1H-NMR and FTIR based Metabolomics approach and UV-Vis, HPTLC and TLC Chromatography Fingerprinting. Research Journal of Pharmacognosy and Phytochemistry. 2023; 15(3): 191-7. http://doi.org/10.52711/0975-4385.2023.00029

35. Corradini C, Cavazza A, Bignardi C. High-performance anion-exchange chromatography coupled with pulsed electrochemical detection as a powerful tool to evaluate carbohydrates of food interest: principles and applications. International Journal of Carbohydrate Chemistry. 2012; 2012. https://doi.org/10.1155/2012/487564

36. Ryu J, Donghyung Y, Byung-Hoo Lee, Suyong Lee. Linkage structure analysis of barley and oat β-glucans by high-performance anion exchange chromatography. Food Science and Biotechnology February 2009 18(1):271-274

37. You J. Determination of β-glucan in health food by high-performance liquid chromatography. Journal of Food Safety and Quality. 2018; 9(1): 34-8.

38. Al-Saffar AZ, Hadi NA, Khalaf HM. Antitumor activity of β-glucan extracted from Pleurotus eryngii. Indian Journal of Forensic Medicine and Toxicology. 2020; Jul 1; 14(3):2493.

39. Allaith SA, Abdel-aziz ME, Thabit ZA, Altemimi AB, Abd El-Ghany K, Giuffrè AM, Al-Manhel AJ, Ebrahim HS, Mohamed RM, Abedelmaksoud TG. Screening and molecular identification of lactic acid bacteria producing β-glucan in boza and cider. Fermentation. 2022; Jul 25; 8(8): 350. https://doi.org/10.3390/fermentation8080350.

40. Khan H. Analytical Method Development in Pharmaceutical Research: Steps involved in HPLC Method Development. Asian Journal of Pharmaceutical Research. 2017; 7(3): 2037. https://doi.org/10.5958/2231-5691.2017.00031.4

41. Yuan H, He Y, Zhang H, Ma X. Ultrasound-assisted enzymatic hydrolysis of yeast β-glucan catalyzed by β-glucanase: Chemical and microstructural analysis. Ultrasonics Sonochemistry. 2022; May 1; 86: 106012. https://doi.org/10.1016/j.ultsonch.2022.106012.

42. Chopade SS, Gaikwad E, Jadhav A, Patil N, Payghan S. Formulation and Evaluation of Fast Disintegrating Tenoxicam Tablets and The Comparison with Marketed Product. 49. https://doi.org/10.5958/2231-5659.2020.00046.6

43. Li Q, Liu J, Zhai H, Zhang Z, Xie R, Xiao F, Zeng X, Zhang Y, Li Z, Pan Z. Extraction and characterization of waxy and normal barley β-glucans and their effects on waxy and normal barley starch pasting and degradation properties and mash filtration rate. Carbohydrate Polymers. 2023; Feb 15; 302: 120405. https://doi.org/10.1016/j.carbpol.2022.120405.

44. Patel B, Rai A, Raut H, Khandhar A, Khunt N. Synthesis of zinc nanoparticle using peppermint leaves and evaluation of zinc nanoparticle by UV, SEM, and XRDS. Research Journal of Pharmacognosy and Phytochemistry. 2022; 14(4): 247-51. https://doi.org/10.52711/0975-4385.2022.00043

45. Kukkar R, Kukkar MR, Shukla SH, Saluja AK. Preparation and Structural Characterization of Kajjali: An Ayurvedic formulation. Research Journal of Pharmacognosy and Phytochemistry. 2012; 4(1): 33-8.

46. Gayatri LA, Amol S, Indrajeet S. Development and validation of a stability-indicating RP-HPLC method for the determination of sitagliptin phosphate and simvastatin in the presence of their degradation products in bulk and binary mixture. Asian Journal of Research in Pharmaceutical Science. 2016; 6(3): 191-7. https://doi.org/10.5958/2231-5659.2016.00026.6

Received on 15.01.2024 Revised on 13.04.2024

Accepted on 28.06.2024 Published on 20.01.2025

Available online from January 27, 2025

Research J. Pharmacy and Technology. 2025;18(1):143-151.

DOI: 10.52711/0974-360X.2025.00022

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