Aspergillus neger SFN-416으로부터 생산한 Xylanase II의 분리정제 및 특성

성찬기,이상원,박석규,손봉수,Sung, Chan-Ki,Lee, Sang-Won,Park, Seok-Kyu,Shon, Bong-Soo 한국미생물 · 생명공학회 1996 한국미생물·생명공학회지 Vol.24 No.6

Xylanase (EC 3.2.1.8) was purified approximately 4.3 fold from Aspergillus niger SFN-416 by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel ion exchange chromatography. Molecular weight of the enzyme was approximately 42,000 daltons. The optimum pH and temperature of the enzyme activity were 5.5 and 50$\circ$C, respectively. The enzyme activity was enhanced by Fe$^{2+}$, and inhibited by Hg$^{2+}$. The activity was decreased by addition of methanol, ethanol, isopropanol and 1-butanol at a concentration of 10%(v/v).

Cellulase를 생산하는 Aspergillus niger MAN - 831과 Aspergillus wentii MAW - 538의 원형질체 형성 및 융합

박석규(Seok-Kyu Park),이상원(Sang-Won Lee),문일식(Il-Shik Moon),손봉수(Bong-Soo Shon),강성구(Seong-Koo Kang) 한국식품영양과학회 1995 한국식품영양과학회지 Vol.24 No.6

섬유성 biomass의 효율적 이용을 위하여 CMCase, avicelase 분비력이 높은 균주(A. wentii MAW-598)와 β-glucosidase(A. niger MAN-831) 분비력이 우수한 균주의 포자를 이용한 원형질체 형성ㆍ재생조건 및 융합을 실시하였다. 원형질체 형성 및 재생의 최적조건은 5일 성숙한 포자를 30㎍/㎖ 농도의 2-DG로 4시간 전배양하여 0.7M KCl 삼투압 안정제, novozyme(7.5㎎/㎖)+driselase(2.5㎎/㎖) 혼합효소로 5시간 반응시키는 것이 효과적이었다. 원형질체 융합 조건은 CaCl₂ 15mM, 25% PEG 4000, pH 6~7, 36℃에서 25~30분간 처리하였을 때 효과적이었으며, 융합빈도는 7.94×10(^-4)로 나타났다. FA-208 융합주는 친주에 비하여 각각 1.5, 1.3, 1.2배 효소활성의 증가를 나타내었다. For the effective utilization of cellulosic biomass, conidial protoplast fusion between Aspergillus niger MAN-831(β-glucosidase) and A. wentii MAW-538(CMCase and avicelase), which produced potently cellulolytic enzymes was carried out. Optimal conditions for formation and regeneration of protoplast were conidiospore age-5 days, 2-DG-30㎍/㎖, preincubation time-4 hours, osmotic stabilizer-0.7M KCl, novozyme(7.5㎎/㎖)+driselase(2.5㎎/㎖) and reaction time of enzyme-5 hours. Optimal conditions for protoplast fusion were obtained by treatment of protoplasts with 15mM CaCl₂ and 25% polyethylene glycol 4000(pH 6~7) as fusogenic agent at 36℃ for 25~30 minutes. The frequency was then 7.94×10^(-4). CMCase, avicelase and β-glucosidase activity of fusant F-208 strain was 1.5, 1.3, 1.2 times higher than those of parental strains, respectively.

Aspergillus awamori와 Zymomonas mobilis로 구성된 혼합고정화 배양계의 최적 조건

이상원(Sang-Won Lee),박석규(Seok-Kyu Park),손봉수(Bong-Soo Shon),최수철(Soo-Cheul Choi),서권일(Kwon-Il Seo),성낙계(Nack-Kie Sung),김홍출(Hong-Chul Kim) 한국식품영양과학회 1995 한국식품영양과학회지 Vol.24 No.5

Gel bead의 효율적인 이용을 목적으로 산소요구성이 전혀 다른 두 균주로 구성된 혼합고정화 배양계의 개발을 시도하였다. 호기성 균으로서 A. awamori, 혐기성균으로서 Z. mobilis를 사용하여 생전분으로 부터 에탄올 생산을 행한 결과는 다음과 같다. 두 균주의 최적 혼합비율은 A. awamori 1.25×10^9 spore/L-gel, Z. mobilis 0.5g cell/L-gel이었다. 배양이 완료된 후의 gel bead 내의 균주 분포는 각각 혐기부와 호기부, 즉 gel bead의 표면부와 중심부로 생육장소가 구분되어 있었다. A-Z계의 배양에서는 에탄올의 수율이 낮았고, 균사가 gel bead로 부터 노출되었다. 배양 개시 후 36시간째에 배양기의 면전을 특수 제조한 check valve가 부착된 실리콘 plug로 교환하여 혐기적 배양을 행한 A-Z 36계에서는 gel bead로 부터 균사의 누출이 상당히 억제되었고, pH가 4.3을 유지하면서, 에탄올의 수율이 대조구 보다 약 2배 높게 나타났다. Co-immobilized mixed culture system(A-Z system) composed of two different oxygen-demanding strains, aerobic(Aspergillus awamori) and anaerobic(Zymomonas mobilis) strains, in a Ca-alginate gel beads was developed to increase ethanol production from raw starch as a carbon source. Optimal mixture ratio of A. awamori and Z. mobilis was 1.25×10^9 pores/L-gel and 0.5g cells/L-gel, respectively. After 120 hours of cultivation, gel beads distinguished oxygen-rich surface for A. awamorifrom oxygen-deficient central part for Z. mobilis. At A-Z culture system, yield of ethanol on glucose, Y_(p/s)=0.18, was very low and there was high leakage of cells from surface of gel beads. At A-Z 36 culture system with changing silicon check valve for cotton plug at 36 hours in A-Z culture system, there was no cell leakage from gel beads, pH was maintained at around 4.3 during cultivation, and yield of ethanol on glucose, Y_(p/s)=0.36, showed 2 times higher than that of control culture system (cotton plug culture).

Xanthomonas sp. EPS-1의 Exopolysaccharide생산을 위한 무기염류의 영향

박석규,손봉수,이상원,서권일,김홍출 순천대학교 기초과학연구소 1995 基礎科學硏究誌 Vol.6 No.-

For the production of high viscosity exopolysaccharide from Xanthomonas sp. EPS-1 which isolated and identified from soils, optimum concentration of inorganic salts were investigated : 0.2% KH_2PO_4, 0.05% MgSO_4·7H_2O, 0.3% NaCl, 0.005% CaCO_3, 0.007% FeSO_4·7H_2O, 0.005% MnSO_4·7H_2O. When EPS-1 strain was cultivated at 30℃ for 72 hours in a jar fermentor (impeller speed 300 rpm, aeration rate 3 vvm, 2 liter), polymer content, product rate and yield(Y_p/s) was 18.4g/ℓ, 0.26g/ℓ/h and 37%, respectively.

균체외 다당류를 생산하는 균주의 분리 및 배양조건

박석규,손봉수,문주석 순천대학교 기초과학연구소 1993 基礎科學硏究誌 Vol.4 No.-

For the screening of a highly viscous exopolysaccharide, a bacterium strain was isolated from soils and identified as Bacillus sp. SFN-83 accroding to the morphological, cultural and physiological properties. The optimum concentration of carbon and nitrogen sources were 1% sucrose and 0.3% ammonium vanadate. At the condition, exopolysaccharide production and product yield were 6.73g/l and 67.3%, respectively.

백일주 양조중 맛성분의 변화

박석규,박필숙,손봉수,문주석 순천대학교 기초과학연구소 1993 基礎科學硏究誌 Vol.4 No.-

Changes in taste components of Backilju, an old Korean cleraed rice wine, were investigated. During brewing, pH and titratable acidity of mash were decreased or increased in the range of 3.41→3.81 and 8.62→6.74, respectively. The final product of Backilju filtrate, fermented for 100 days, contained 0.36% reducing sugar, 2.68% total sugar, 606.6 ㎎% total acid, 2.5 ㎎% volatile acid and 22.9㎎ solid/ml extract. Major volatile components analized by GC/MS methods were 3-methyl-1-butanol acetate, 4-decanol and dimethyl disulfide. Calcium(20.26ppm) was the most abundant, followed by magnesium(3.19ppm) and zinc(1.87ppm). Nucleotides and their related compounds were inosine(0.12→0.44 mM/ml), ADP(0.16→0.33 mM/ml) and hypoxanthine(0.33→0.04 mM/ml). Compared with other Korean traditional Yakju, Backilju contained more total acid and mineral but less sugar and volatile acid.

Aspergillus niger SFN-416으로부터 생산한 Xylanase 2의 분리정제 및 특성

성찬기,이상원,박석규,손봉수 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

Aspergillus niger SFN-416으로부터 생성된 xylanase Ⅱ를 분리·정제하여 특성을 조사하였다. Aspergillus niger SFN-416의 배양액을 ethanol(70%) 침전, (NH_4)_2SO_4(30~90%) 침전, Sephadex G-100 chromatography 및 DEAE-Sephacel chromatography 등의 정제과정을 거친 결과, specific activity가 38.2 units/ mg protein으로 4.3배 정제되었고, 정제효소의 최적 활성온도는 50℃였다. 최적 pH는 5.5이었고, pH 4.5~6.0 사이에서 비교적 안정하였다. 또한 금속이온에 대한 효소의 활성은 대부분 억제를 보였고, 특히 Hg^2+에서는 3.7%로 강하게 효소활성이 저해됨을 보였고, Fe^2+에서는 100%의 상대활성을 나타내었다. 정제 효소의 분자량은 SDS-PAGE에 의하여 42,000 daltons이었으며, 유기용매에 대한 활성은 10%의 methanol, ethanol, isopropanol 및 1-butanol에 대하여 모두 낮은 활성을 나타었다. Xylanase (EC 3.2.1.8) was purified approximately 4,3 fold from Aspergillus niger SFN-416 by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel ion exchange chromatography. Molecular weight of the enzyme was approximately 42,000 daltons. The optimum pH and temperature of the enzyme activity were 5.5 and 50℃, respectively. The enzyme activity was enhanced by Fe^2+, and inhibited by Hg^2+. The activity was decreased by addition of methanol, ethanol, isopropanol and 1-butanol at a concentration of 10%(v/v).

Aspergillus awamori와 Zymomonas mobilis로 구성된 고정화혼합 배양계의 에탄올 생산에 미치는 Triton, PVA 및 PEG의 영향

이상원,서권일,박석규,손봉수,김홍출,성찬기 순천대학교 기초과학연구소 1995 基礎科學硏究誌 Vol.6 No.-

The effects of triton-X 100, PVA and PEG on the ethanol production and raw starch hydrolysis by co-immobilized A. awamori and Z. mobilis (A-Z system) were investigated. When gel beads containing A. awamori(1.25 × 10 exp (9) spores/ℓ - gel) and Z. mobilis(0.5g cell/ℓ - gel) were cultured in 2% raw starch solution at 30℃ for 120 hours, ethanol concentration, ethanol yield (Y_p/s) and pH of culture broth were 2.8g/ℓ, 0.16 and 3.8~4.0, respectively. Ethanol production and starch hydrolysis by addition of PVA was efficient, compared to that of PEG and ethanol concentration in culture broth with 0.2% PVA was 3.56g/ℓ. At A-Z 36 culture system with changing silicon check valve for cotton plug at 36 hours in A-Z culture system, ethanol yield on glucose, Y_p/s = 0.36, showed 2 times higher than that of control culture system(cotton plug culture). Ethanol production and productivity by cultivating 85g gel beads at 30℃ in a 1ℓ jar fermentor(220rpm) of medium 500㎖ were 32g/ℓ and 1.8g/ℓ/day, respectively. After 12 hours of cultivation, gel beads were distinguished oxygen-rich surface for A. awamori from oxygen-deficient central part for Z. mobilis.