• Applied Chemistry for Engineering
• /
• v.22 no.3
• /
• pp.336-339
• /
• 2011

Algae is an abundant and potential fermentation substrate. The enzymatic hydrolysis of algae was investigated by pre-treating an alkaline hydrogen peroxide with commercial cellulase and viscozyme. Algae used in this study was the Ulva pertusa. The evaluated response was the yield of released glucose after the enzymatic hydrolysis. Alkaline hydrogen peroxide containing mixtures of 1 wt% hydrogen peroxide and 1~1.75 wt% sodium hydroxide was also used. The results show that the highest glucose conversion was obtained for Ulva pertusa using 5 wt% hydrogen peroxide at $60^{\circ}C$ for 3 h. The required amount of enzymes after the pre-treatment with alkaline hydrogen peroxide were reduced by far compared to that of untreated Ulva pertusa. Also, the amount of glucose that is released during the enzymatic hydrolysis was increased.

• Journal of Korea Technical Association of The Pulp and Paper Industry
• /
• v.33 no.3
• /
• pp.37-43
• /
• 2001

The bleach efficiency between PY and YP bleaching of pulps, which were deinked in neutral (cellulase) and alkaline (NaOH, NaOH/${Na_2}{SiO_3}$) ranges, and were treated on kneader, was compared in this study. The brightness of pulps (84.8~88.4% ISO) which were deinked in alkaline or neutral ranges increased up to 88.4%~88.8% ISO after PY or YP bleaching. There was no difference in the brightness between PY and YP bleaching. And the dirt count of bleached pulps was similar each other at the range of 17~38 ppm. The tensile index of bleached pulps was slightly higher in PY bleaching than those in YP bleaching. While with the treatment of kneading, the brightness and dirt count decreased remarkably compared to those of no-kneading. However, the sharp difference in the brightnesses or in dirt count of bleached pulps decreased after bleaching. The brightness of bleached pulps at YP stages was only numberically higher than those at PY stages without regard to the treatment of kneading. Therefore, in the case of the neutral deinkiing process, the conversion of PY bleaching stages to YP bleaching stages was profitable to water loop system, and was harmless in the aspect of the bleach efficiency.

• Korean Journal of Agricultural Science
• /
• v.25 no.1
• /
• pp.131-137
• /
• 1998

The cellulase components were partially purified from the culture filtrate of the alkalophilic bacterium Pseudomonas sp. AC-711 and its enzymatic properties were characterized. The specific activity of the purified major enzyme component was 3.5 units/mg protein as carboxymethyl cellulase and the yield was 23% of the total activity of the culture broth. The molecular weight of the component was 46,000 and the Km and Vmax on CMC were determined as $15.4mg\;mL^{-1}$ and $4.17{\mu}moles\;mL^{-1}\;min^{-1}$, respectively. The enzyme was stable at the temperatures below $60^{\circ}C$ and at the pH range of 4.0~11.0, and the optimal temperature and pH were $60^{\circ}C$ and pH 8.0, respectively. The enzyme activity was not significantly affected by the common surfactants (concentration: 0.05%) such as ${\alpha}$-olefin sulfonate, linear alkylbenzene sulfonate, sodium dodecyl sulfonate, hexadecyltrimethylammonium bromide and Tween 80. The enzyme was activated by the metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Co^{2+}$, whereas inhibited by $Hg^{2+}$ and $Zn^{2+}$. The enzyme exhibited relatively high activity toward amorphous CMC as compared with crystalline substrates such as filter paper and avicel.

• Animal Bioscience
• /
• v.34 no.5
• /
• pp.867-879
• /
• 2021

Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.10
• /
• pp.1427-1437
• /
• 2014

Enzymatic hydrolysis of lignocellulosic biomass into fermentable sugars is a key step in the conversion of agricultural by-products to biofuels and value-added chemicals. Utilization of a robust microorganism for on-site production of biomass-degrading enzymes has gained increasing interest as an economical approach for supplying enzymes to biorefinery processes. In this study, production of multi-polysaccharide-degrading enzymes from Aspergillus aculeatus BCC199 by solid-state fermentation was improved through the statistical design approach. Among the operational parameters, yeast extract and soybean meal as well as the nonionic surfactant Tween 20 and initial pH were found as key parameters for maximizing production of cellulolytic and hemicellulolytic enzymes. Under the optimized condition, the production of FPase, endoglucanase, ${\beta}$-glucosidase, xylanase, and ${\beta}$-xylosidase was achieved at 23, 663, 88, 1,633, and 90 units/g of dry substrate, respectively. The multi-enzyme extract was highly efficient in the saccharification of alkaline-pretreated rice straw, corn cob, and corn stover. In comparison with commercial cellulase preparations, the BCC199 enzyme mixture was able to produce remarkable yields of glucose and xylose, as it contained higher relative activities of ${\beta}$-glucosidase and core hemicellulases (xylanase and ${\beta}$-xylosidase). These results suggested that the crude enzyme extract from A. aculeatus BCC199 possesses balanced cellulolytic and xylanolytic activities required for the efficient saccharification of lignocellulosic biomass feedstocks, and supplementation of external ${\beta}$-glucosidase or xylanase was dispensable. The work thus demonstrates the high potential of A. aculeatus BCC199 as a promising producer of lignocellulose-degrading enzymes for the biomass conversion industry.

• Korean Journal of Food Science and Technology
• /
• v.27 no.3
• /
• pp.370-374
• /
• 1995

By exposing the complex enzyme solution to alkaline condition, it was possible to remove the protease activity selectively without inactivation of soybean cell wall degrading activity of the crude enzyme complex produced by Aspergillus niger CF-34. Optimum reaction conditions were as follow. pH was $9.0{\pm}0.1$, temperature was $20^{\circ}C$ and reaction time was 30 min with gentle stirring. Over 90% of protease activity could be eliminated while the activities of pectinase, polygalacturonase, xylanase, carboxymethyl cellulase and soybean cell wall degrading enzyme were maintained to $80{\sim}100%$. Through alkali treatment, it was discovered that the quality and organoleptic properties of soy protein produced by this enzymes were improved because the hydrolysis of protein and formation of bitter peptide were decreased.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.5
• /
• pp.690-695
• /
• 2016

This study was performed to evaluate the effect of alkaline treatment on Styela clava tunic (SCT). Considerable damage to the surface of alkali-treated SCT was observed by scanning electron microscopy (SEM) in a concentration-dependent manner upon alkaline treatment. The amount of crystalline region in SCT gradually increased with increasing NaOH concentration, which was analyzed by X-ray diffraction and thermogravimetric analysis. The initial enzymatic reaction of Celluclast toward SCT was elevated by treatment with NaOH up to 1.0 N concentration due to disruption of the SCT surface by promoting binding of enzymes with SCT. However, in the late stage of the enzyme reaction, hydrolysis rate decreased with elevation of NaOH concentration, thereby increasing the amount of non-reacted residuals. This result was due to the increase in the crystalline regions in SCT.

• Applied Biological Chemistry
• /
• v.51 no.4
• /
• pp.299-304
• /
• 2008

A ${\beta}$-1,4-endoglucanase gene from Bacillus subtilis H12 was cloned into Escherichia coli JM109 (pBC8) and sequenced. The endoglucanase gene with an insert DNA of 2.5 kb possessed an open reading frame of 1,500 bp encoding a mature protein of 499 amino acids with a calculated molecular mass of 55 kDa. The deduced amino acid sequence showed similarity to those of the known neutral cellulase genes of B. subtilis PAP115 (99.2%) and BSE616 (97.8%), as well as the alkaline gene of Bacillus sp. N4 (55.1%). The endoglucanase activity expressed by E. coli (pBC8) was localized in the periplasmic fraction (80%) and the cytoplasmic fraction (20%). An endoglucanase was purified from the periplasmic fraction by performing gel filtration and anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 31 kDa by SDS-PAGE, and the maximum activity occurred at pH 7 and $40^{\circ}C$. The enzyme easily hydrolyzed soluble substrates such as carboxymethyl cellulose and barely ${\beta}$-glucan, whereas the sigmacell and xylan, the known insoluble substrates, were not entirely hydrolyzed.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.11
• /
• pp.1525-1535
• /
• 2014

The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

• Korean Journal of Soil Science and Fertilizer
• /
• v.29 no.2
• /
• pp.173-180
• /
• 1996

Effects of inoculation with phosphate-solubilizing microorganisms, Pseudomonas putida and Aspergillus niger, were studied in both acidic red-yellow and alkaline calcareous soils cropped with pimiento. In red-yellow soil after cultivation, the amounts of soil available phosphorus on non-fertilizer and fertilizer plots inoculated with Aspergillus niger, and on rice straw plot inoculated with Pseudomonas putida and Aspergillus niger were significantly higher than uninoculation treatments, but there were no differences in calcareous soil. With inoculation of either Pseudomonas putida or Aspergillus niger, increase in phosphorous uptake by pimiento cultivated in calcareous soil was detected on non-fertilizer, and fertilizer plots except rice straw plot. Although there were no significant differences in soil cellulase activities among treatments, the activity was the highest on rice straw plot in red-yellow soil. The phosphatase activities in red-yellow soil were increased by the inoculation with Aspergillus niger only, and the activity in calcareous soil was improved by the inoculation with either Pseudomonas putida or Aspergillus niger.