Bacillus subtilis 분리균의 Mannanase와 Xylanase 생산성과 효소특성
Production and Properties of Mannanase and Xylanase by a Bacillus subtilis Isolate
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초록
새만금 갯벌로부터 xylan과 locust bean gum (LBG)을 분해하는 미생물을 분리하여 그 생화학적 특성과 16S rRNA 서열을 조사한 결과 Bacillus subtilis로 확인되었다. 분리균 YB-30은 konjac이나 LBG가 존재한 상태에서 배양할 경우 mannanase의 생산성이 급격하게 증가되며, 밀기울이 존재하에서는 xylanase의 생산성이 증가되였다. Konjac (3.5%) 과 밀기울(1%)이 동시에 첨가된 배지에서 균의 성장이 정지기에 진입하였을 때 mannanase와 xylanase의 생산성이 최고 수준에 도달하였다. 배양상등액의 mannanase와 xylanase는 60℃와 pH 6.0, 55℃와 pH 5.5에서 각각 최대 활성을 보였다. 고온에서는 두 효소 모두 안정하지는 않았으며 xylanase가 mannanase보다 안정성이 낮았다. 밀기울은 쌀기울보다많은 양의 arabinoxylan을 함유하고 있으므로 배양상등액으로 분해하였을 때 밀기울로부터 생성되는 환원당의 양이 더 많은 것으로 확인되었으며, 이로 보아 B. subtilis YB-30에의해 생산되는 xylanase와 mannanase는 사료첨가용 효소로 활용 가능성이 높다고 여겨진다.
Abstract
A bacterial strain capable of hydrolyzing xylan and locust bean gum (LBG) was isolated from the Saemangeum tideland of Korea. Based on the biochemical properties and the 16S rRNA gene sequence, the isolate YB-30 was identified as Bacillus subtilis. Xylanase productivity was increased effectively when B. subtilis YB-30 was grown in the presence of wheat bran, while mannanase productivity was increased drastically when grown in the presence of konjac or LBG. Particularly, maximum mannanase and xylanase activities were detected in the culture filtrate of media containing 3.5% konjac and 1% wheat bran. Both enzyme productivities reached maximum levels in the stationary growth phase. The culture filtrate exhibited the highest activity at 60℃ and pH 6.0 for mannanase and at 55℃ and pH 5.5 for xylanase, respectively. Both enzymes were not stable at high temperatures and xylanase was less stable than mannanase. In addition, wheat bran was hydrolyzed to liberate reducing sugar to a greater extent than rice bran by the culture filtrate because the wheat bran contained more arabinoxylan than the rice bran. Hence, xylanase and mannanase produced by B. subtilis YB-30 have a potential use as feed additive enzymes.
참고문헌 (23)
- Akhavan Sepahy A, "Costeffective production and optimization of alkaline xylanase by indigenous Bacillus mojavensis AG137 fermented on agricultural Waste", Enzyme Res, 2011, pp.593624~ (2011)[DOI]
- Amore A, "Application of a new xylanase activity from Bacillus amyloliquefaciens XR44A in brewer's spent grain saccharification", J. Chem. Technol. Biotechnol, 90, pp.573~581 (2015)
- Cavaglieri L, "Biocontrol of Bacillus subtilis against Fusarium verticillioides in vitro and at the maize root level", Res. Microbiol, 156, pp.748~754 (2005)
- Chang P, "Cloning and characterization of two thermostable xylanases from an alkaliphilic Bacillus firmus", Biochem. Biophys. Res. Commun, 319, pp.1017~1025 (2004)
- Huang J, "Cloning, sequencing and expression of the xylanase gene from a Bacillus subtilis strain B10 in Escherichia coli", Bioresour. Technol, 97, pp.802~808 (2006)
- Huang JL, "Highlevel production of a cold-active β-mannanase from Bacillus subtilis BS5 and its molecular cloning and expression", Mol. Gen. Mikrobiol. Virusol, 4, pp.14~17 (2012)
- Izydorczyk MS, "Cereal arabinoxylans:advances in structure and physicochemical properties", Carbohydr. Polym, 28, pp.33~48 (1995)
- Jiang Z, "High-level production, purification and characterization of a thermostable β-mannanase from the newly isolated Bacillus subtilis WY34", Carbohydr. Polym, 66, pp.68~96 (2006)
- Jorgensen HA, "Production of ethanol and feed by high dry matter hydrolysis and fermentation of palm kernel press cake", Appl. Biochem. Biotechnol, 161, pp.318~332 (2010)
- Koullas DP, "Correlating the effect of pretreatment on the enzymatic-hydrolysis of straw", Biotechnol. Bioeng, 39, pp.113~116 (1992)
- Li, Yanan, "Gene Cloning, Expression, and Characterization of a Novel β-Mannanasefrom Bacillus circulans CGMCC 1416", Journal of Microbiology and Biotechnology, 18(1), pp.160~166 (2008), 한국미생물·생명공학회
- Nagar S, "Production and optimization of cellulase-free, alkali-stable xylanase by Bacillus pumilus SV-85S in submerged fermentation", J. Ind. Microbiol, 37, pp.71~83 (2010)
- Prakash P, "Production of alkaliphilic, halotolerent, thermostable cellulase free xylanase by Bacillus halodurans PPKS-2 using agro waste: single step purification and characterization", World J. Microbiol. Biotechnol, 28, pp.183~192 (2012)
- Ralet MC, "Influence of extrusion-cooking on the physicochemical properties of wheat bran", J. Cereal Sci, 11, pp.793~812 (1990)
- Saleem M, "Characterization of a thermostable and alkaline xylanase from Bacillus sp. and its bleaching impact on wheat straw pulp", World J. Microbiol. Biotechnol, 28, pp.513~522 (2012)
- Subramaniyan S, "Biotechnology of microbial xylanases: enzymology, molecular biology, and application", Crit. Rev. Biotechnol, 22, pp.33~64 (2002)
- van Dyk JS, "Identification of endoglucanases, xylanases, pectinases and mannanases in the multi-enzyme complex of Bacillus licheniformis SVD1", Enzyme Microb. Technol, 47, pp.112~118 (2010)
- Vijayalaxmi S, "Production of extremely alkaliphilic, halotolerent, detergent, and thermostable mannanase by the free and immobilized cells of Bacillus halodurans PPKS-2. Purification and characterization", Appl. Biochem. Biotechnol, 171, pp.382~395 (2013)
- Wongputtisin P, "Screening and selection of Bacillus spp. for fermented corticate soybean meal production", J. Appl. Microbiol, 113, pp.798~806 (2012)
- Xie X, "Isolation and characterization of wheat bran starch", Food Res. Int, 41, pp.882~887 (2008)
- 윤기홍, "Bacillus amyloliquefaciens 분리균의 Mannanase 생산성과 효소특성", 미생물학회지, 50(2), pp.158~163 (2014), 한국미생물학회[DOI]
- 윤기홍, "Characterization of the Bacillus subtilis WL-3 Mannanase from a Recombinant Escherichia coli", The Journal of Microbiology, 46(3), pp.344~349 (2008), 한국미생물학회
- Zangiabadi H, "The effect of a β-mannanasebased enzyme on growth performance and humoral immune response of broiler chickens fed diets containing graded levels of whole dates", Trop. Anim. Health Prod, 42, pp.1209~1217 (2010)