Cellulase Extract Research Articles

Cellulases Production byAspergillus unguis in Solid State Fermentation and Enzymatic Hydrolysis of Dilute Acid-Treated Groundnut Fodder.

Groundnut fodder was utilized as a bioresource for the production of cellulases through solid state fermentation (SSF). Aspergillus unguis was initially grown on modified groundnut fodder for cellulase production and the fodder was hydrolyzed by the crude cellulase extract into fermentable hydrolyzate. The highest titer of Filter paperase (FPase), Carboxymethyl cellulase (CMCase), β-glucosidase, and protein content were found to be 11.4, 11.5, 33.3 U/g, and 136.6mg/g of substrate, respectively. Treatment of groundnut fodder with dilute acid and supplementation of mineral salts, and yeast extract (5% w/v) enhanced cellulase production. The hydrolysis of modified groundnut fodder treated with Tween-80 using the crude cellulase extract resulted in increased production of reducing sugars (40%) at 50°C compared with control. In conclusion, the addition of Tween-80 at pH 4.8 increased a twofold increase in the yield of reducing sugar.

Characterization of Major Cell-Wall-Degrading Enzymes Secreted by Diaporthe spp. Isolate Z1-1N Causing Postharvest Fruit Rot in Kiwifruit in China.

Pathogen-induced fruit decay is a significant threat to the kiwifruit industry, leading to considerable economic losses annually. The cell-wall-degrading enzymes (CWDEs) secreted by these pathogens are crucial for penetrating the cell wall and accessing nutrients. Among them, Diaporthe species are recognized as major causal agents of soft rot in kiwifruit, yet their pathogenic mechanisms are not well understood. In this study, we explored the production of various CWDEs secreted by Diaporthe Z1-1N, including polygalacturonase (PG), polymethylgalacturonase (PMG), polygalacturonic acid transeliminase (PGTE), pectin methyltranseliminase (PMTE), endoglucanase (Cx), and β-glucosidase (β-glu), both in liquid cultures and within infected kiwifruit tissues. Our findings revealed significant activities of two pectinases (PG and PMG) and cellulases (Cx and β-glu) in the infected tissues. In contrast, very low levels of PMTE and PGTE activities were observed under the same conditions. When orange pectin served as the carbon source, PG and PMG showed notable activities, while PMTE and PGTE remained inactive. Moreover, the activities of Cx and β-glu significantly decreased by more than 63 times in the liquid medium with carboxymethyl cellulose (CMC) as the carbon source compared to their levels in infected kiwifruit. A further analysis indicated that the necrotic lesions produced by pectinase extracts were larger than those produced by cellulase extracts. Notably, four enzymes-PG, PMG, Cx, and β-glu-exhibited high activities on the third or fourth day post-infection with Diaporthe Z1-1N. These results suggest that Diaporthe Z1-1N secretes a range of CWDEs that contribute to kiwifruit decay by enhancing the activities of PG, PMG, Cx, and β-glu. This study sheds light on the pathogenicity of Diaporthe in kiwifruit and highlights the importance of these enzymes in the decay process.

Optimization of extraction of crude polysaccharide from Cassia seed via ultrasonic assisted cellulase and study on its antioxidant activity

In order to improve the current low extraction rate of cassia polysaccharide and improve its extraction efficiency, the ultrasonic wave assisted cellulase extraction method of cassia polysaccharide is first proposed and optimized. Orthogonal test is conducted to analyze the influence of six factors related to cellulase and ultrasonic wave on the extraction of cassia polysaccharide, and at the same time to explore the antioxidant activity of the extracted polysaccharide in vitro. The results show that the main and secondary factors affecting the extraction rate of cassia polysaccharide are pH > ultrasonic power > enzyme dosage > enzymatic hydrolysis temperature > enzymatic hydrolysis time > ultrasonic time. The optimal conditions are: enzyme dosage: 1.6%, enzymatic hydrolysis time: 80 min, enzymatic hydrolysis temperature: 52 ℃, enzymatic hydrolysis pH: 6.4, ultrasonic time: 30 min, ultrasonic power: 276 W, the polysaccharide yield under the optimal conditions is 12.29%, which is 5.31% and 14.43% higher than the single enzyme extraction method and ultrasonic extraction method reported in the literature, respectively, indicating that the composite extraction method is superior to the single extraction method. The antioxidant activity analysis shows that the half inhibitory concentrations of cassia polysaccharide against DPPH·, ABTS·+, ·OH, ·O2- free radicals are 0.894 mg/mL, 1.175 mg/mL, 1.908 mg/mL and 7.601 mg/mL, respectively, which shows good antioxidant activity in vitro. Therefore, ultrasonic assisted cellulase extraction of crude polysaccharide from cassia seed is a new efficient and rapid extraction method.

Impact of Six Extraction Methods on Molecular Composition and Antioxidant Activity of Polysaccharides from Young Hulless Barley Leaves.

Young hulless barley leaves are gaining recognition for potential health benefits, and the method of extracting polysaccharides from them is critical for potential food industry applications. This study delves into a comparative analysis of six distinct fiber extraction techniques: hot water extraction; high-pressure steam extraction; alkaline extraction; xylanase extraction; cellulase extraction; and combined xylanase and cellulase extraction. This analysis included a thorough comparison of polysaccharide-monosaccharide composition, structural properties, antioxidant activities (DPPH, ABTS, and FRAP), and rheological properties among fibers extracted using these methods. The results underscore that the combined enzymatic extraction method yielded the highest extraction yield (22.63%), while the rest of the methods yielded reasonable yields (~20%), except for hot water extraction (4.11%). Monosaccharide composition exhibited divergence across methods; alkaline extraction yielded a high abundance of xylose residues, whereas the three enzymatic methods demonstrated elevated galactose components. The extracted crude polysaccharides exhibited relatively low molecular weights, ranging from 5.919 × 104 Da to 3.773 × 105 Da across different extraction methods. Regarding antioxidant activities, alkaline extraction yielded the highest value in the ABTS assay, whereas enzymatically extracted polysaccharides, despite higher yield, demonstrated lower antioxidant capacity. In addition, enzymatically extracted polysaccharides exerted stronger shear thinning behavior and higher initial viscosity.

Ultrasonic/cellulase-assisted extraction of polysaccharide from Garcinia mangostana rinds and its carboxymethylated derivative

Response surface methodology was selected to explore the ultrasonic-assisted cellulase extraction conditions of Garcinia mangostana rind polysaccharides (GMRPs), and the optimum values of each condition were as follows: ratio of raw material to liquid of 1:50 g/mL, ultrasonic time of 40 min, enzyme concentration of 4 %, and ultrasonic power of 179 W. Based on the above conditions, the average extraction rate of GMRPs was 15.56 %. GMRPs were modified by carboxymethylation, and the relationship between the amount of chloroacetic acid and the substitution degree of carboxymethylated derivative was compared. Based on the results of single factor experiment, it was shown that the amount of chloroacetic acid significantly affected the degree of substitution of derivative products. The above research provides some valuable theoretical references for the preparation of GMRPs and its carboxymethylation products.

Multi-frequency ultrasound-assisted cellulase extraction of protein from mulberry leaf: Kinetic, thermodynamic, and structural properties

The effects of different extraction methods (traditional extraction, ultrasound extraction, cellulase extraction, and ultrasound-assisted cellulase extraction) on the yield of mulberry leaf protein (MLP) were investigated, and the results revealed that multi-frequency ultrasound-assisted cellulase extraction was the most efficient extraction method. The mechanism of the synergistic extraction method used to efficiently extract protein from mulberry leave was investigated, focusing on the kinetics and thermodynamics of the enzymatic process. The results revealed that kinetic parameters KM decreased by 14.07% and kA increased by 5.02%, and the thermodynamic parameters Ea, ΔH, and ΔS decreased by 44.81%, 48.41%, and 21.12 %, respectively, following the process of multi-frequency ultrasound (MFU) pretreatment. The spectral analysis with fluorescence spectra manifested that ultrasound exposed hydrophobic groups and induced molecular unfolding of MLP. Atomic force microscope showed that ultrasound decreased particle size while increasing flexibility of MLP. The effect of ultrasound increases the binding frequency of cellulase and substrates, resulting in greater affinity between the two and promoting the solubilization of MLP. This study provides a theoretical basis to improve the application prospects of MLP.

Effect of enzyme – assisted extraction on total polyphenol content and antioxidant capacity from fresh tea leaves (Camellia sinensis)

AbstractTea leaves contain high concentration of polyphenols, known as bioactive compounds. In order to increase the extraction yield of polyphenol and the antioxidant capacity of the extract from tea leaves, the combination of cellulase, pectinase and protease in enzyme‐assisted extraction was investigated. By using fractional factorial experimental design to study the two‐enzyme assisted extraction of cellulase and pectinase, three factors including solvent/material ratio, pH and hydrolysis time showed significant effects on the extract properties. The highest total polyphenol content obtained adopting the favorable conditions was 185.9 mg GAE/g of dry material at solvent and material ratio of 5.9, hydrolysis pH of 4.4 and hydrolysis time of 36 minutes. In addition, the antioxidant capacity of the extract was determined at 175.9 mg TE/g of dry material at those optimum experimental conditions. In our test of three‐enzyme (cellulase – pectinase – protease) assisted extraction, the extracts were spray‐dried to observe green tea powder. Results indicated that the TPC, AC and the color sensory quality of the green tea powder decreased compared to the two‐enzyme (cellulase – pectinase) assisted extraction.

Effects of Four Extraction Methods on Structure and In Vitro Fermentation Characteristics of Soluble Dietary Fiber from Rape Bee Pollen.

In this study, soluble dietary fibers (SDFs) were extracted from rape bee pollen using four methods including acid extraction (AC), alkali extraction (AL), cellulase extraction (CL) and complex enzyme extraction (CE). The effects of different extraction methods on the structure of SDFs and in vitro fermentation characteristics were further investigated. The results showed that the four extraction methods significantly affected the monosaccharide composition molar ratio, molecular weight, surface microstructure and phenolic compounds content, but showed little effect on the typical functional groups and crystal structure. In addition, all SDFs decreased the Firmicutes/Bacteroidota ratio, promoted the growth of beneficial bacteria such as Bacteroides, Parabacteroides and Phascolarctobacterium, inhibited the growth of pathogenic bacteria such as Escherichia-Shigella, and increased the total short-chain fatty acids (SCFAs) concentrations by 1.63-2.45 times, suggesting that the bee pollen SDFs had a positive regulation on gut microbiota. Notably, the SDF obtained by CE exhibited the largest molecular weight, a relatively loose structure, higher extraction yield and phenolic compounds content and the highest SCFA concentration. Overall, our results indicated that CE was an appropriate extraction method of high-quality bee pollen SDF.

Extraction Optimization, Characterization and Biological Activities of Polysaccharide Extracts from Nymphaea hybrid.

In this study, polysaccharide-rich Nymphaea hybrid extracts (NHE) were obtained using the ultrasound-assisted cellulase extraction (UCE) method optimized by response surface methodology (RSM). The structural properties and thermal stability of NHE were characterized by Fourier-transform infrared (FT-IR), high-performance liquid chromatography (HPLC) and thermogravimetry-derivative thermogravimetry (TG-DTG) analysis, respectively. Moreover, the bioactivities of NHE, including the antioxidant, anti-inflammatory, whitening and scratch healing activities were evaluated by different in vitro assays. NHE conveyed a good ability to scavenge against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals and inhibit the hyaluronidase activity. NHE can effectively protect the HaCaT cells against oxidative damage by inhibiting the intracellular reactive oxygen species (ROS) production in the H2O2 stimulation assays and promoting the proliferation and migration in the scratch assays. In addition, NHE was proven to inhibit melanin production in B16 cells. Collectively, the above results seem to be the evidence needed to promote the potential of NHE to be regarded as a new functional raw material in the cosmetics or food industries.

Physicochemical, functional and biological properties of soluble dietary fibers obtained from Rosa roxburghii Tratt pomace using different extraction methods

Rosa roxburghii fruit pomace is an abundant, but underutilized, byproduct from the juice industry. In this study, the effects of different extraction methods on physicochemical, functional and biological properties of water soluble dietary fibers (SDFs) extracted from R. roxburghii Tratt pomace were compared. The SDFs, named RWSDF, RXSDF and RCSDF, were extracted from R. roxburghii Tratt pomace by water extraction, xylanase extraction, and cellulase extraction methods. Results indicated that RWSDF, RXSDF and RCSDF varied in chemical and monosaccharide compositions. The structural results indicated that RWSDF, RXSDF and RCSDF exhibited similar spectral characteristics of polysaccharides. Compared with RWSDF, RXSDF and RCSDF exhibited higher crystallinity, higher thermal stability, and more porous structures. Furthermore, RXSDF and RCSDF showed higher water-holding capacity, oil-holding capacity, water solubility, nitrite ion absorption capacity, antioxidant activity, and α-glucosidase inhibitory activity. These findings suggest that RRTP may be a potential functional food and a promising natural source of dietary fiber.

CELLULASE OF ENDOPHYTIC Bacillus SP. FROM Glycyrrhiza uralensis F. AND ITS APPLICATION FOR EXTRACTION OF GLYCYRRHIZIC ACID

Glycyrrhizic acid is the main component of the medicinal plant Glycyrrhiza uralensis Fisch. It is widely used as a sweetener and an effective active ingredient with multiple physiological functions. Endophytes are microorganisms that coexist with plants and can produce cellulase. This cellulase enzyme can be used to overcome dissolution barriers of plant active ingredients by degrading plant cell wall. In the present study, a cellulase-producing strain with high cellulase activity was isolated from fresh Glycyrrhiza uralensis Fisch, and identified using the Congo red staining method and the DNS method. Glycyrrhizic acid yield was determined by the HPLC method. A highly reactive cellulase-producing strain, with a high extraction capacity of glycyrrhizic acid, was obtained. The strain was named GG-3, and bioinformatic analysis showed that it was a Bacillus sp. Findings obtained after optimization of the enzyme production and glycyrrhizic acid extraction process showed that glycyrrhizic acid yield increased by 32.52% and 31.35% after extraction with GG-3 enzyme, compared with the use of the traditional extraction method and commercial cellulase extraction method, respectively.

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

In line with the depletion of petroleum reserves from fossils, humans have forced people to look for alternative renewable energy sources. Nipah plant stems are organic material that contains a lot of lignocellulose. Cellulose in lignocellulose can be hydrolyzed to glucose to produce bioethanol. Problems arise because hydrolysis using pure cellulase requires relatively high costs, so it is necessary to find an alternative source of cheap cellulase. One of the cheap sources of cellulase is goat rumen fluid. Until now, there is not much information regarding the reaction kinetics and cellulase characteristics of goat rumen fluid. The purpose of this study was to isolate crude cellulase extract, determine the KM and Vmax values ​​and determine the optimum temperature and pH. The research was carried out at the Unsoed Agricultural Technology Laboratory. The samples used were seven rumen fluids. The study began with isolation followed by enzymatic kinetics studies. Fractionation was carried out by adding ammonium sulfate salt (50, 60,70, and 80 %), and centrifugation at 7,000 rpm at 4 oC for 15 minutes. Crude cellulase extract with the highest enzyme activity value was used for the study of enzymatic reaction kinetics and characterization. The crude cellulase resistance test to temperature was carried out at 45, 55 and 65 oC and the pH was carried out at pH 4, 5, 6, 7 and 8. The KM and Vmax values ​were determined by measuring the activity of crude cellulase extract at various concentrations of CMC (1, 1 ,5, 2, 2.5, and 3 %). The results showed that crude cellulase extract had an average specific activity of 1.7356 IU/mg. The highest enzyme activity was 0.9854 IU/ml. The optimum crude cellulase extract was at pH 6 with an activity of 1.1015 IU/ml and a temperature of 60 oC with an activity of 0.7829 IU/ml. At a CMC concentration of 2.5% crude cellulase extract had an activity of 0.3179 IU/ml with a Vmax value of 0.0045 IU/ml and a KM of 0.0252 %.

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

In line with the depletion of petroleum reserves from fossils, humans have forced people to look for alternative renewable energy sources. Nipah plant stems are organic material that contains a lot of lignocellulose. Cellulose in lignocellulose can be hydrolyzed to glucose to produce bioethanol. Problems arise because hydrolysis using pure cellulase requires relatively high costs, so it is necessary to find an alternative source of cheap cellulase. One of the cheap sources of cellulase is goat rumen fluid. Until now, there is not much information regarding the reaction kinetics and cellulase characteristics of goat rumen fluid. The purpose of this study was to isolate crude cellulase extract, determine the KM and Vmax values ​​and determine the optimum temperature and pH. The research was carried out at the Unsoed Agricultural Technology Laboratory. The samples used were seven rumen fluids. The study began with isolation followed by enzymatic kinetics studies. Fractionation was carried out by adding ammonium sulfate salt (50, 60,70, and 80 %), and centrifugation at 7,000 rpm at 4 oC for 15 minutes. Crude cellulase extract with the highest enzyme activity value was used for the study of enzymatic reaction kinetics and characterization. The crude cellulase resistance test to temperature was carried out at 45, 55 and 65 oC and the pH was carried out at pH 4, 5, 6, 7 and 8. The KM and Vmax values ​were determined by measuring the activity of crude cellulase extract at various concentrations of CMC (1, 1 ,5, 2, 2.5, and 3 %). The results showed that crude cellulase extract had an average specific activity of 1.7356 IU/mg. The highest enzyme activity was 0.9854 IU/ml. The optimum crude cellulase extract was at pH 6 with an activity of 1.1015 IU/ml and a temperature of 60 oC with an activity of 0.7829 IU/ml. At a CMC concentration of 2.5% crude cellulase extract had an activity of 0.3179 IU/ml with a Vmax value of 0.0045 IU/ml and a KM of 0.0252 %.

Extraction and characterization of cellulase from forest and compost soil fungal isolates for the application of straw degradation

Cellulases are complex hydrolytic enzymes working synergistically on the hydrolysis of cellulolytic materials for the production of simple sugars. These enzymes have tremendous environmental, industrial and agricultural applications including enhancement of the degradability of lignocellulosic materials for cattle’s feed. The aim of this study was to isolate fungus and extract cellulose enzyme from forest and compost soil samples and examine the extent to which these enzymes enhance the degradability of finger millet and oat straw for making palatable cattle’s feed. Accordingly, a total of 53 fungal isolates were isolated from forest and compost soil. On 1% CMC media resulted 40% were cellulolytic fungal species. Six were selected based on their clear zone. These isolates belonged to the genera Trichoderma, Aspergillus and Penicillium based on their morphological characteristics. Cultivation of fungal isolates for cellulase production using submerged fermentation and Solid-state fermentation was undertaken through make the variation on their levels of different growth conditions such as temperature, pH, minerals and substrates. The results showed that the highest cellulase production i.e CMCase 83.12 ± 3.18 U/dL and FPase 44.51± 0.391FPU/dL were obtained from FSI6 in FMS supplemented SSF at 280C and a pH of 6.

Optimization of Ultrasound-Assisted Cellulase Extraction from Nymphaea hybrid Flower and Biological Activities: Antioxidant Activity, Protective Effect against ROS Oxidative Damage in HaCaT Cells and Inhibition of Melanin Production in B16 Cells.

In this study, ultrasonic-assisted cellulase extraction (UCE) was applied to extract flavonoids and polyphenols from the Nymphaea hybrid flower. The extraction conditions were optimized using the response surface method (RSM) coupled with a Box-Behnken design. The crude extract of Nymphaea hybrid (NHE) was further purified using AB-8 macroporous resins, and the purified extract (NHEP) was characterized by FTIR and HPLC. In vitro activity determination by chemical method showed that NHEP displayed strong free radical scavenging abilities against the DPPH and ABTS radicals, good reduction power, and hyaluronidase inhibition. The cell viability by CCK-8 assays showed that NHEP had no significant cytotoxicity for B16 and HaCaT cells when the concentration was below 100 μg/mL and 120 μg/mL, respectively. NHEP with a concentration of 20–160 μg/mL can more effectively reduce the ROS level in H2O2 damaged HaCaT cells compared with 10 μg/mL of VC. The 40 μg/mL of NHEP had similar activity against intracellular melanin production in the B16 melanoma cells compared with 20 μg/mL Kojic acid. Good activities of antioxidation, whitening and protective effect against H2O2-induced oxidative damage promote the potential for NHEP as a functional raw material in the field of cosmetics and medicine.

Harnessing the potential use of cellulolytic Klebsiella oxytoca (M21WG) and Klebsiella sp. (Z6WG) isolated from the guts of termites (Isoptera)

BackgroundFor many years, denim-heavy quality cotton twill colored with indigo colors and with a well-worn/faded look has held a lot of appeal. Machine damage, drainage system blockage, and other issues come with the conventional usage of pumice stones for “stone-washing” denims. In view of the abovementioned information, a range of works has been done to investigate the economic prospects of bacterial cellulase enzymes for use in industrial processes, including biopolishing in the textile sector. Ethiopia has excellent termite diversity to isolate bacterial gut-associated cellulose enzymes for biostoning applications. The main purpose of this study was, therfore, to decipher how to isolate and characterize cellulase enzymes from termite (Isoptera) gut bacteria with the intention of employing it for biostoning of textiles.PurposeTo use cellulolytic enzymes of Klebsiella oxytoca (M21WG) and Klebsiella sp. (Z6WG) isolated from termite guts in biostoning of textiles and improving garment quality.MethodsCellulase enzyme-producing bacteria were isolated and screened from the guts of worker termites sampled from Meki and Zeway termite mounds in the Central Rift Valley region of Ethiopia. Bacterial screening, biochemical, morphological, and 16S rRNA sequence identification techniques were employed to characterize the bacterial strains. In addition, the production, optimization, and purification of the associated cellulase enzymes were employed, and the potential application of the enzymes for biostoning of a textile was demonstrated.ResultThe isolated M21WG was found to be 99% identical to the Klebsiella oxytoca (MT104573.1) strain, while the isolated Z6WG showed 97.3% identity to the Klebsiella sp. strain (MN629242.1). At an ideal pH of 7, a temperature of 37 °C, a 72-h incubation time, and a substrate concentration of 1.5% carboxymethylcellulose sodium, the maximum activity of the crude cellulase extract from these bacteria was assessed. These bacteria produced cellulase enzymes that were moderately efficient. Consequently, it was determined that the cellulase enzymes were effective for biostoning of denim cloth.ConclusionIt was determined that Klebsiella oxytoca (M21WG) and Klebsiella sp. (Z6WG) could be used as a doorway to better understand harnessing the use of these cellulase-producing bacteria from termite (Isoptera) guts. In this study, it was also attempted to assess the effectiveness of the two bacterial isolates in biostoning in anticipation of their potential application in the textile realm.

Saccharification of acid-alkali pretreated sugarcane bagasse using immobilized enzymes from Phomopsis stipata.

The online version contains supplementary material available at 10.1007/s13205-021-03101-2.

Optimization of Ultrasonic Cellulase-Assisted Extraction and Antioxidant Activity of Natural Polyphenols from Passion Fruit.

In this paper, ultrasonic cellulase extraction (UCE) was applied to extract polyphenols from passion fruit. The extraction conditions for total phenol content (TPC) and antioxidant activity were optimized using response surface methodology (RSM) coupled with a Box-Behnken design (BBD). The results showed that the liquid-to-solid ratio (X2) was the most significant single factor and had a positive effect on all responses. The ANOVA analysis indicated quadratic models fitted well as TPC with R2 = 0.903, DPPH scavenging activity with R2 = 0.979, and ABTS scavenging activity with R2 = 0.981. The optimal extraction parameters of passion fruit were as follows: pH value of 5 at 30 °C for extraction temperature, 50:1 (w/v) liquid-to-solid ratio with extraction time for 47 min, the experimental values were found matched with those predicted. Infrared spectroscopy suggested that the extract contained the structure of polyphenols. Furthermore, three main polyphenols were identified and quantified by HPLC. The results showed the content of phenolic compounds and antioxidant activity of the optimized UCE were 1.5~2 times higher than that determined by the single extraction method and the Soxhlet extraction method, which indicates UCE is a competitive and effective extraction technique for natural passion fruit polyphenols.

Optimised cellulase production from fungal co-culture of Trichoderma reesei NCIM 1186 and Penicillium citrinum NCIM 768 under solid state fermentation

Microbial cellulases find applications in many industries and constitute a significant share of the world’s industrial enzyme market. In order to improve the cost function of the cellulose producing processes with enhanced yield and novel activities, superior bioprocesses are formulated these days. Intending to produce higher titre of cellulase enzyme, present study deals with fungal co-culture and to investigate its feasibility of producing higher cellulase titre via solid state fermentation (SSF). Various process parameters were optimized for induced cellulase production from the co-culture of Trichoderma reesei NCIM 1186 and Penicillium citrinum NCIM 768 in SSF. Among different low cost and easily available agricultural lignocellulosic waste, wheat bran was found to be the most suitable substrate. The highest cellulase yield of 6.71 FPU/gds is obtained on 6th day of fermentation using steam pre-treated wheat bran with 70 % moisture content when inoculated with 106 spores of mixed suspension at 30 °C and pH 5. A further possibility to use whole fermented matter for saccharification instead of extracted cellulase was also investigated with the aim of abolishing an extraction step. Akin sugar yield of 6.45 mg/mL and 6.58 mg/mL was obtained at 72 h of hydrolysis of steam pre-treated wheat bran, using cellulase extract and whole fermented matter, respectively. Fungal co-culture of T. reesei and P. citrinum in SSF results in the highest cellulase titre using low cost agricultural waste as a substrate, which can be further used for successful saccharification in the form of whole fermented matter or as extracted enzyme.

Effects of Cellulase and Pectinase Extraction on the Antioxidant and Antitumor Activities of Licorice Residue Polysaccharide

Objective:To investigate the effects of different enzyme extraction methods on the antioxidant and antitumor properties of licorice residue polysaccharide and to provide evidence for the effective reuse of licorice. Methods:The licorice residue polysaccharides were extracted by cellulase and pectinase under optimized conditions respectively. The yield,IR spectra,antioxidant activity and antitumor activity were investigated and compared. Results:The yield of licorice residue polysaccharide extracted by pectinase and cellulase was 8.43% and 10.71%,respectively. The IR spectra confirmed the skeletal modes of the pyranose ring in licorice residue polysaccharide. The IC50 values of DPPH,hydroxyl and ABTS radical scavenging activity of polysaccharide extracted by pectinase and cellulase were 0.00243,0.305,0.0403 mg/mL and 0.226,0.0238,0.0401 mg/mL,respectively. Antitumor results showed that the inhibition rate of cellulase extraction polysaccharide was higher than pectinase extraction polysaccharide in range of 0.00250~0.125 mg/mL. Conclusion:Cellulase extraction polysaccharide had high yield,better hydroxyl radical scavenging activity and antitumor property. Pectinase extraction polysaccharide had better DPPH free radical scavenging activity. The scavenging ability of ABTS free radical of them was same.