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흑두로 제조한 청국에서 분리된 Bacillus subtillus BB-1으로부터 혈전용해효소 유전자 크로닝 및 특성규명
이영훈,이성호,전주미,김홍출,조용운,박기훈,최영주,갈상완 Plant molecular biology and biotechnology research 2005 Plant molecular biology and biotechnology research Vol.2005 No.
A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla ⅠDNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript ⅡSK(-) and showed the fibrinolytic activity to bacterial cells. The Cla ⅠDNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla ⅠDNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and 35℃, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.
송영민,진상근,갈상완,이성대,김성찬,정맹화 진주산업대학교 농업기술연구소 2001 農業技術硏究所報 Vol.14 No.-
To estimate the essential and nonessential amino acid compositions as added moisture levels in manufacturing fermentation feedstuffs, the three kinds of experimental conditions, T1, T2, T3 were made with rice bran(90kg), leaf mold(10kg) according to added moisture 40kg, 60kg, and 80kg, respectively. In T1 and T2 cases, the contents of threonine, phenylalanine, histidine, aspartic acid, glutamic acid, cystine, arginine increased significantly during fermentation, however, in T3 case, the thereonine content increased significantly after 9 hours fermentation but decreased at 12 hours fermentation(P<0.05). In T1 and T3 cases, the contents of valine, leucine and arginine increased significantly after 9 hours fermentation and decreased at 12 hours fermentation(P<0.05). Methionine increased for three hours fermentation and then did not increase in content in the T1 and the T3. In T2 case, methionine level increased gradually for whole fermentation time. In T3 case, phenylalanine, histidine, aspartic acid, glutamic acid and cystine increased significantly after 9 hours fermentation, but decreased at 12 hours fermentation(P<0.05). In T1, T2 and T3, isoleucine, lysine, serine, proline, glycine, alanine and tyrosine increased significantly during fermentation time(P<0.05).
전통발효식품으로부터 Chitin 분해 미생물의 분리 및 특성 규명
고보경,최인순,이상현,임채오,이성호,갈상완,최영주 Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-
A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophaeus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide (NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37℃ and pH 7.8~8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.
Sang Wan Gal(갈상완),Yea Hwang Moon(문여황),Soo Jeong Cho(조수정) 한국생명과학회 2011 생명과학회지 Vol.21 No.12
본 실험에서는 새송이버섯 수확 후 배지 발효산물 첨가급여가 한우 거세우의 성장과 도체성적에 미치는 영향을 조사하기 위하여 비육기(17-24개월령) 한우 거세우 32두를 공시하여 30개월령까지 사양시험을 수행하였다. 공시동물은 30%의 새송이버섯 수확 후 배지 발효산물(F-SMS)이 첨가된 TMR 사료를 급여한 처리구와 시판 배합사료와 볏짚을 분리 급여한 대조구로 나누어 처리구별로 각각 16두씩 배치하였으며 한 우리(5×6 m)당 4두씩 수용하였다. 30%의 F-SMS가 첨가된 TMR 사료의 영양 가치를 분석한 결과, 30%의 F-SMS가 첨가된 TMR 사료는 볏짚보다는 조단백질(11.67%)과 TDN(72.11%) 함량이 높게 나타났으나 시판배합사료와는 큰 차이가 없었다(p<0.05). 시험기간 동안 사료섭취량은 30%의 F-SMS가 첨가된 TMR 사료 급여구가 대조구보다 유의적으로 많았으나(p<0.05) 공시동물의 증체량에서는 처리구간에 큰 차이가 없었다. 육질등급에서는 30%의 F-SMS가 첨가된 TMR 사료 처리구(100%)의 1등급 이상 출현율이 대조구(87.5%)보다는 높게 나타났으나 육량등급에 있어서는 처리구별로 큰 차이가 없었다(p<0.05). 이상의 결과로 볼 때, Bacillus amyloliquefaciens CS47와 Saccharomyces cerevisiae로 발효시켜 기호성과 저장성이 개선된 새송이버섯 수확 후 배지는 한우 거세우의 원료사료로 사용 가능한 것으로 사료된다. This study was carried out to investigate the feeding effects of the dietary supplementation of fermented spent mushroom (Pleurotus eryngii) substrates with Bacillus amyloliquefaciens CS47 and Saccharomyces cerevisiae (F-SMS) on growth performance and carcass characteristics of Hanwoo steers. Thirty two Hanwoo steers were allocated into two feeding groups and assigned equally to two dietary treatments; Control (commercial formula feed for Hanwoo steers and rice straw) and TMR including 30% F-SMS. The nutritional values of TMR including 30% F-SMS was higher crude protein (11.67%) and TDN (72.11%) than rice straw, but not significantly different from commercial formula feed (p<0.05). Feed intake was significantly greater in the TMR including 30% F-SMS than the control (p<0.05), but body weight gain and carcass grades were not influenced by the experimental diets. Based on this study, fermented spent mushroom (Pleurotus eryngii) substrate with Bacillus amyloliquefaciens CS47 and Saccharomyces cerevisiae is may be used as an ingredient of feed in TMR for Hanwoo steer.
Sang-Sang Park,Suk-Hee Sung,Young-Bae Ryu,Young-Un Cho,Ki-Hoon Park,Sang-Wan Gal 한국버섯학회 2008 한국버섯학회지 Vol.6 No.2
This study was carried out to investigate the growth inhibition of Propionibacterium acnes by mycelial culture broth of Paecilomyces japonica in the Mulberry leaf extract. The results are as following. The growth inhibition effect of P. japonica mycelial culture broth against P. acnes in various concentration of Mulberry leaf extract was the most effective in 3% Mulberry leaf extract. The inhibition effect of P. japonica mycelial culture broth against P. acnes in Mulberry leaf extract according to the culture time was the moust effective for 25 days cultivation. As the treating volume of the mycelial culture broth was increased, the growth inhibition effect against P. acnes was increased as well. The growth inhibition effect of mycelial culture broth against P. acnes according to the time of heat treatment was active by 45min at 100℃, while it was inactive at more than 60min. Taken together P. japonica mycelial culture in the Mulbarry leaf extract has a possibility to be an element of skincare cosmetics regulating the acnes. This means that the enzyme may be used for health-care such as thrombosis without any hamful behavour in the blood vessel.
Molecular Cloning and Characterization of 58 kDa Chitinase Gene from Serratia marcescens KCTC 2172
Gal Sang Wan,Lee S. W.,Choi Y. J. The Korean Society for Biotechnology and Bioengine 2002 Biotechnology and Bioprocess Engineering Vol.7 No.1
A chitinase gene (pCHi58) encoding a 58 kDa chitinase was isolated from the Serratia marcescens KCTC 2172 cosmid library. The chitinase gene consisted of a 1686 bp open reading frame that encoded 562 amino acids. Escherichia coil harboring the pChi58 gene secreted a 58 kDa chitinase into the culture supernatant. The 58 kDa chitinase was purified using a chitin affinity column and mono-S column. A nucleotide and N-terminal amino acid sequence analysis showed that the 58 kDa chitinase had a leader peptide consisting of 23 amino acids which was cleaved prior to the 24th alanine. The 58 KDa chitinase exhibited a $98\%$ similarity to that of S. marcescens OMB 1466 in its nuclotide sequence. The chitinolytic patterns of the 58 kDa chitinase released N,N'-diacetyl chitobiose (NAG2) as the major hydrolysis end-product with a trace amount of N-acetylglucosamine. When a 4-methylumbellyferyl-N-acetylglucosamin monomer, dimmer, and tetramer were used as substrates, the 58 kDa chitinase did not digest the 4-Mu-NAG monomer $(analogue\;of\;NAG_2)$, thereby indicating that the 58 kDa chitinase was likely an endochitinase. The optimum reaction temperature and pH of the enzyme were $50^{\circ}C$ and 5.0, respectively.
PURIFICATION AND CHARACTERIZATION OF CHITINASE ISOFORMS FROM S.marcescens ATCC27117
Gal, Sang-Wan,Kim, Cha-Young,Bae, Chang-Gyu,Kang, Chang-Ho,Bahk, Jeong-Dong,Lee, Sang-Yeol,Cho, Moo-Je Gyeongsang National University Chinju, Korea 1992 Plant molecular biology and biotechnology research Vol.1992 No.
길상완(Sang Wan Gal),조수정(Soo Jeong Cho) 한국생명과학회 2011 생명과학회지 Vol.21 No.10
버섯 생산 후 발생되는 부산물인 팽이버섯 수확 후 배지로부터 7종의 고온성 균주를 분리하였으며 이 중 곰팡이독소를 생성하는 Asp. flavus와 Asp. ochraceous에 대한 항균활성이 높으면서 xylanase와 cellulase 생성능이 우수한 균주를 최종 선발하여 UJ03으로 명명하였다. Bacillus ID kit와 VITEK 2 system를 이용하여 분리균 UJ03의 생리적·생화학적 특성을 조사한 결과 분리균 UJ03은 Bacillus 속과 유사한 특징을 나타내었으며 16S rDNA 염기 서열 분석 결과에서는 B. amyloliquefaciens와 98.9%의 상동성을 나타내었다. 이와 같은 결과를 종합하여 분리균 UJ03은 Bacillus sp. UJ03으로 동정되었으며 분리균 UJ03이 생성하는 항균물질은 TLC와 HPLC 분석에서 Bacillus 속 균주가 생성하는 펩타이드성 항균물질인 iturin A와 유사한 특성을 나타내었다. Spent mushroom substrate (SMS) is a by-product remained after a crop of mushrooms. About seven thermophilic strains were isolated from SMS (Flammulina velvtipes). Among them, one isolate, designated UJ03, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ03 produced cellulase and xylanase as extracellular hydrolases. The strain UJ03 was identified as a member of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ03 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ03 was classified within the genus Bacillus, for which the name Bacillus sp. UJ03 is proposed. The antifungal compound from Bacillus sp. UJ03 was similar to lipopeptide iturin A of Bacillus sp.