• 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.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.5
• /
• pp.802-810
• /
• 2007

Fibrobacter succinogenes, a Gram-negative, anaerobic ruminal bacterium is a major fibre digesting species in the rumen. It intensively degrades plant cell walls by an erosion type of mechanism, burrowing its way through the complex matrix of cellulose and hemicellulose with the release of digestible and undigested cell wall fragments. The enzymes involved in this process include a combination of glucanases, xylanases, arabinofuranosidase(s) and esterases. The genome of the bacterium has been sequenced and this has revealed in excess of 100 putative glycosyl hydrolase, pectate lyase and carbohydrate esterase genes, which is greater than the numbers reported present in other major cellulolytic organisms for which genomes have been sequenced. Modelling of the amino acid sequences of two glycanases, CedA and EGB, by reference to crystallized homologs has enabled prediction of the major features of their tertiary structures. Two dimensional gel electrophoresis in conjunction with mass spectroscopy has permitted the documentation of proteins over expressed in F. succinogenes grown on cellulose, and analysis of the cell surfaces of mutant strains unable to bind to cellulose has enabled the identification of candidate proteins with roles in adhesion to the plant cell wall substrate, the precursor to cellulose biodegradation.

• Mycobiology
• /
• v.42 no.2
• /
• pp.181-184
• /
• 2014

To the best of our knowledge, this is the first report on thermophilic fungi isolated in Korea. Three species of thermophiles were isolated from compost and were identified as Myriococcum thermophilum, Thermoascus aurantiacus, and Thermomyces lanuginosus. They can grow at temperatures above $50^{\circ}C$ and produce high levels of cellulolytic and xylanolytic enzymes at high temperatures. Notably, the considerable thermostability of the endo-glucanase produced by T. aurantiacus has made the fungus an attractive source of industrial enzymes.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.7
• /
• pp.965-970
• /
• 2016

This study was conducted to determine the effects of freezing and supplementation of molasses (M), lactic acid bacteria (LAB) and LAB+enzyme mixture on chemical and nutritional composition of sunflower silage (SF). Sunflower crops were harvested (at about $29.2%{\pm}1.2%$ dry matter) and half of fresh sunflower was ensiled alone and half was frozen (F) at $-20^{\circ}C$ for 7 days. Silage additives were admixed into frozen SF material. All samples were ensiled in glass jars with six replicates for 90 days. The treatments were as follows: i) positive control (non-frozen and no additives, NF), ii) negative control (frozen, no additives, F), iii) F+5% molasses (FM), iv) F+LAB (1.5 g/tons, Lactobacillus plantarum and Enterococcus faecium, FLAB); v) F+LAB+enzyme (2 g/tons Lactobacillus plantarum and Enterococcus faecium and cellulase and amylase enzymes, FLEN). Freezing silage increased dry matter, crude ash, neutral detergent fiber, and acid detergent lignin. The organic matter, total digestible nutrient, non-fiber carbohydrate, metabolizable energy and in vitro dry matter digestibility were negatively influenced by freezing treatments (p<0.05). In conclusion, freezing sunflower plants prior to ensiling may negatively affect silage quality, while molasses supplementation improved some quality traits of frozen silage. Lactic acid bacteria and LAB+enzyme inoculations did not effectively compensate the negative impacts of freezing on sunflower silage.

• Korean Journal of Medicinal Crop Science
• /
• v.21 no.2
• /
• pp.97-104
• /
• 2013

The objective of this study is to investigate the optimal condition for macerating enzymatic extraction process that leads to the highest yield and the largest extracted amount of bio-active contents from Rubus coreanus Miq. fruit. The optimal extraction conditions were found as the following: The initial amount of the water added to the fruit was 20 ~ 30% by weight. The mixing ratio used for the macerating enzyme was 4 : 1 : 2 (w : w : w) for cellulase:pectinase:amylogucosidase, and the amount of the macerating enzyme added was 2% by weight. The extraction process was done at a temperature of $45{\sim}50^{\circ}C$ for 10 hours. The extraction yields on Rubus coreanus Miq. fruit by macerating enzymatic extraction process was increased by 84.3% compared to that of hot-water extraction process. The amounts of organic acids and vitamin found in the extract were also higher. The amount of polyphenol and anthocyanin contents in the extract were 185% and 257% of those from hot-water extraction, respectively. These results suggest that macerating enzymatic extraction is an effective method to boost extraction yield and to increase the amount of extraction of bio-active contents from Rubus coreanus Miq. fruit.

• KSBB Journal
• /
• v.29 no.4
• /
• pp.297-302
• /
• 2014

The autoclaving assisted by an irradiation pretreatment method was developed without toxic chemicals to produce fermentable sugars for their conversion to bioethanol. In the first step, electron beam irradiation (EBI) of rice straw was performed at various doses. The electron beam-irradiated rice straw was then autoclaved with DI water at $120^{\circ}C$ for 1 h. A total sugar yield of 81% was obtained from 300 kGy electron beam-irradiated rice straw after 72 h of enzymatic hydrolysis by Cellulase 1.5L (70 FPU/mL) and Novozyme-188 (40 CbU/mL). Also, the removal of hemicellulose and lignin was 32.0% and 32.5%, respectively. This result indicates that the environmentally-friendly pretreatment method of rice straw by an electron beam irradiation could be applied for bioethanol production in plant.

• Journal of the Korean Society of Clothing and Textiles
• /
• v.37 no.3
• /
• pp.348-356
• /
• 2013

Denim washing is processed with different washing techniques such as stone washing, chemical washing, sand washing, and bio washing. Cellulase bio washing can meet environmental regulations that enhance and rectify problems associated with traditional decolorization techniques; however, stone washing needs to be added to the processing because it produces a low decolorization effect. There is also the problem of additional strength reduction. To prevent these problems, a new enzyme for bio washing is required. This study examines the optimum laccase treatment conditions on denim and evaluated the characteristics of laccase-treated denims to establish a database of eco-friendly new decolorization process on denim using a new laccase enzyme. The results show that the optimum conditions of laccase on denim are a pH of 4.0, $30^{\circ}C$, 7% (o.w.f.), and 6 hours in 10 mM of buffer concentration. UV absorbance and HPLC identified isatin coexist with anthranilic acid in solution after laccase treatment on denim. Results of the surface color, the surface morphology and the tensile strength indicate that laccase treatment shows an excellent decolorization effect without fiber damage. The wet cleaning fastness and the perspiration fastness also improved.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.3
• /
• pp.340-345
• /
• 1999

A novel simultaneous saccharification and fermentation (SSF) process from the microcrystalline cellulose to ethanol was developed by using $\delta$-integrated recombinant cellulolytic Saccharomyces cerevisiae L2612$L2612\deltaGC$, which can utilize cellulose as carbon and energy sources. The optimum amount of enzymes needed for the efficient conversion of cellulose to ethanol at $30^{\circ}C$ was determined with commercial cellulolytic enzymes. By fed-batch cultivation, the heterologous cellulolytic enzymes were accumulated up to 42.67% of the total cellulase and 29% of the $\beta$-glucosidase needed for the efficient SSF process. When this $\delta$-integrated recombinant yeast was applied to the successive SSF step for ethanol production, 20.35 g/l of ethanol was produced after 12 h from 50 g/l of microcrystalline cellulose. By using this novel SSF process, a considerable amount of commercial enzymes was reduced.

• Journal of Applied Biological Chemistry
• /
• v.50 no.4
• /
• pp.202-210
• /
• 2007

A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

• Journal of Korea Technical Association of The Pulp and Paper Industry
• /
• v.32 no.4
• /
• pp.58-65
• /
• 2000

Use of a pectinase during preparing handmade papers from bast fiber of paper mulberry(Broussonetia kazinoki Sieb.) was investigated in order to decrease cooking chemicals and environmental pollution. For this purpose, four kinds of commercial pectinases, Rapidase LIQ(RLP), Rapidase Press(RP), Rapidase C80L Max(RCM) and Pectinase SS Kyowa(PSK) were used. And the durability of handmade papers before and after pectinase treatment was determined. RP and PSK had higher pectinase activity ad lower cellulase activity. The bast fiber was not defibered when pectinase was used. In order to increase the efficiency of enzymes, the bast fiber were treated ammonium oxalate(AO) or $K_2CO_3$under mild conditions. The AO pretreatment with those produced by $K_2CO_3$. The RP treated pulps after mild $K_2CO_3$cooking of the bast fiber were defibrated more easily than untreated pulp. The handmade paper prepared with the RP treated pulps after mild $K_2CO_3$cooking has good strength properties such as breaking length and folding endurance. Also, it has higher durability on heat aging, though its brightness was slightly lower than that of untreated paper.