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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제22권 제3호
- 발행연도
- 2012.1
- 수록면
- 331 - 338 (8page)
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초록· 키워드
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A novel gene coding for an endo-β-1,4-mannanase (manA)from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The β-mannanase showed an identity of 90.2-92.9% (≤95%)with the corresponding amino acid sequences from B.
subtilis strains deposited in GenBank. The purified β-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDITOF mass spectrometry. The recombinant β-mannanase had an optimum temperature of 45oC and optimum pH of 6.5. The enzyme was stable at temperatures up to 50oC (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions (Hg2+, Pb2+, and Co2+)substantially inhibited the recombinant β-mannanase.
The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol,ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v)or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the β-mannanase expressed in P.
pastoris could potentially be used as an additive in the feed for monogastric animals.
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참고문헌 (21)
참고문헌 신청
Benech, R. / 2007 / Recombinant expression, characterization, and pulp prebleaching property of a Phanerochaete chrysosporium endo-beta-1,4-mannanase / Enzyme Microb. Technol
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem
Dhawan, S. / 2007 / Microbial mannanase: An overview of production and applications / Crit. Rev. Biotechnol 27 : 197 ~ 216
Ethier, N. / 1998 / Gene cloning, DNA sequencing, and expression of thermostable beta-mannanase from Bacillus stearothermophilus / Appl. Environ. Microbiol 64 : 4428 ~ 4432
Hatada, Y. / 2005 / Sequence of the gene for a high-alkaline mannanase from an alkaliphilic Bacillus sp. strain JAMB-750, its expression in Bacillus subtilis and characteriza
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem
Dhawan, S. / 2007 / Microbial mannanase: An overview of production and applications / Crit. Rev. Biotechnol 27 : 197 ~ 216
Ethier, N. / 1998 / Gene cloning, DNA sequencing, and expression of thermostable beta-mannanase from Bacillus stearothermophilus / Appl. Environ. Microbiol 64 : 4428 ~ 4432
Hatada, Y. / 2005 / Sequence of the gene for a high-alkaline mannanase from an alkaliphilic Bacillus sp. strain JAMB-750, its expression in Bacillus subtilis and characteriza
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