OBJECTIVETo investigate the optimal extraction process of polysaccharides from S. fusiforme by enzymatic treatment.

METHODThe optimum extraction conditions were obtained by the experiment with the orthogonal design. The content of polysaccharides of S. fusiforme was determined by spectraphotometry.

RESULTThe amount of enzyme and temperature significantly affected total polysaccharides of S. fusiforme.

CONCLUSIONThe optimum extraction conditions include the addition of 1. 2 x 10 (4) U x 100 g(-1) enzyme into water at pH 4. 5, and the subsequent treatment for 10 min while the temperature is maintained at 45 degrees C.

Cellulase ; metabolism ; Hydrogen-Ion Concentration ; Polysaccharides ; analysis ; isolation & purification ; metabolism ; Sargassum ; chemistry ; Technology, Pharmaceutical ; methods ; Temperature

Cellulose takes nearly 10% (W/W) dry weight of cassava tubers. In this study, the cellulase cost of different ethanol fermentation from cassava cellulose was evaluated. The processes include the direct saccharification and fermentation of original cassava cellulose residues, the direct saccharification and fermentation of pretreated cassava cellulose residues, and the simultaneous co-saccharification and fermentation of cassava starch and cassava cellulose. The results show that the cassava cellulose utilization in the first two processes were low with the enzyme cost of 13 602 and 11 659 RMB Yuan per tone of ethanol, respectively. In the third process, the final ethanol concentration increased from 101.5 g/L to 107.0 g/L when cassava cellulose and cassava starch were saccharified simultaneously. Comparing to the first two processes, the third one demonstrated the lowest enzyme cost at 3 589 RMB Yuan per ton of ethanol, which was less than the ethanol price and no additional equipment and operation cost input were added. The conclusion provided a practical way of cassava cellulose utilization in cassava ethanol industry.
Biotechnology ; economics ; methods ; Cellulase ; economics ; Cellulose ; metabolism ; Cost-Benefit Analysis ; Ethanol ; economics ; metabolism ; Fermentation ; Manihot ; metabolism ; Saccharomyces cerevisiae ; metabolism

High-concentration sugars production from stover is an important perspective technology for the cellulosic ethanol industrialization. Fed-batch process is an effective way to achieve this goal in the fermentation industry. In this study, based on fed-batch process, high-concentration sugars were produced from pretreated corn stover by enzymatic hydrolysis. After being pretreated by the dilute sulphuric acid, the impacts of the ratio of solid raw material to liquid culture, the content of supplementary materials and the refilling time on the saccharification rate were investigated. Results showed that the initial ratio of solid raw material to liquid culture was 20% (W/V) and the initial concentrations of enzymes for xylanase, cellulose and pectinase were 220 U, 6 FPU, and 50 U per gram of substrates, respectively. After 24 hours and 48 hours, 8% pretreated corn stovers were added respectively together with the additions of xylanase (20 U) and cellulose (2 FPU) per gram of substrates. After 72 hours, the final concentration of reducing sugar was increased to 138.5 g/L from 48.5 g/L of the non fed-batch process. The rate of enzyme hydrolysis of the raw material was 62.5% of the thoretical value in the fed-batch process. This study demonstrated that the fed-batch process could significantly improve the concentration of reducing sugar.
Batch Cell Culture Techniques ; methods ; Carbohydrates ; analysis ; Cellulase ; metabolism ; Cellulose ; metabolism ; Endo-1,4-beta Xylanases ; metabolism ; Ethanol ; metabolism ; Fermentation ; Hydrolysis ; Plant Stems ; chemistry ; metabolism ; Zea mays ; chemistry

During the bioconversion of lignocellulose to ethanol, the biomass always undergoes pretreatment in order to increase the enzymatic digestibility of cellulose. In present work, we conducted the pretreatment of sugarcane bagasse with aqueous acetic acid for delignification and alkali for deacetylation respectively (Acetoline process) to increase cellulose accessibility for enzymatic hydrolysis. The effects of several factors on the pretreatment effectiveness were investigated. The enzymatic digestibility of pretreated bagasse was further studied. The enzymatic glycan conversion of pretreated solid reached about 80% when it was digested under 7.5% solid consistency with cellulase of 15 FPU/g solid and beta-glucosidase of 10 CBU/g solid for 48 h. Compared with dilute acid pretreatment, Acetoline pretreatment could obtain higher enzymatic glycan conversion. The experimental results indicate that Acetoline is an effective pretreatment method to increase the enzymatic digestibility of sugarcane bagasse.
Acetic Acid ; chemistry ; Biotechnology ; methods ; Cellulase ; metabolism ; Cellulose ; chemistry ; metabolism ; Energy-Generating Resources ; Ethanol ; analysis ; metabolism ; Hydrolysis ; Lignin ; chemistry ; metabolism ; Saccharum

β-Glucosidase activity assays constitute an important indicator for the early diagnosis of neonatal necrotizing enterocolitis and qualitative changes in medicinal plants. The drawbacks of the existing methods are high consumption of both time and reagents, complexity in operation, and requirement of expensive instruments and highly trained personnel. The present study provides a simplified, highly selective, and miniaturized glucometer-based strategy for the detection of β-glucosidase activity. Single-factor experiments showed that optimum β-glucosidase activity was exhibited at 50 °C and pH 5.0 in a citric acid-sodium citrate buffer when reacting with 0.03 g/mL salicin for 30 min. The procedure for detection was simplified without the need of a chromogenic reaction. Validation of the analytical method demonstrated that the accuracy, precision, repeatability, stability, and durability were good. The linear ranges of β-glucosidase in a buffer solution and rat serum were 0.0873-1.5498 U/mL and 0.4076-2.9019 U/mL, respectively. The proposed method was free from interference from β-dextranase, snailase, β-galactosidase, hemicellulase, and glucuronic acid released by baicalin. This demonstrated that the proposed assay had a higher selectivity than the conventional dinitrosalicylic acid (DNS) assay because of the specificity for salicin and unique recognition of glucose by a personal glucose meter. Miniaturization of the method resulted in a microassay for β-glucosidase activity. The easy-to-operate method was successfully used to detect a series of β-glucosidases extracted from bitter almonds and cultured by Aspergillus niger. In addition, the simplified and miniaturized glucometer-based assay has potential application in the point-of-care testing of β-glucosidase in many fields, including medical diagnostics, food safety, and environmental monitoring.
Animals ; Aspergillus niger ; Calibration ; Cellulase/analysis* ; Chemistry, Clinical/methods* ; Dextranase/analysis* ; Enterocolitis, Necrotizing/diagnosis* ; Equipment Design ; Flavonoids/analysis* ; Glucose/analysis* ; Glucuronic Acid/analysis* ; Glucuronidase/analysis* ; Glycoside Hydrolases/analysis* ; Hydrogen-Ion Concentration ; Linear Models ; Multienzyme Complexes/analysis* ; Plants, Medicinal ; Polygalacturonase/analysis* ; Rats ; Reproducibility of Results ; beta-Galactosidase/analysis* ; beta-Glucosidase/analysis*

Genes encoding the cellobiohydrolase enzyme (CBHI), designated as cbhI, were isolated from the basidiomycetes Auricularia fuscosuccinea, Pleurotus giganteus, P. eryngii, P. ostreatus, and P. sajor-caju. Initially, the fungal genomic DNA was extracted using a modified cetyltrimethyl ammonium bromide (CTAB) protocol and used as a DNA template. The cbhI genes were then amplified and cloned using the pGEM-T Easy Vector Systems. The sizes of these PCR amplicons were between 700~800 bp. The DNA sequences obtained were similar showing high identity to the cbhI gene family. These cbhI genes were partial consisting of three coding regions and two introns. The deduced amino acid sequences exhibited significant similarity to those of fungal CBHI enzymes belonging to glycosyl hydrolase family 7.
Amino Acid Sequence ; Base Sequence ; Basidiomycota ; Bromides ; Cellulase ; Cellulose 1,4-beta-Cellobiosidase ; Clinical Coding ; Clone Cells ; Cloning, Organism ; DNA ; Humans ; Introns ; Pleurotus ; Polymerase Chain Reaction ; Quaternary Ammonium Compounds ; Sequence Analysis