Purification and Characterization of Alkaline Pectin Lyase from a Newly Isolated Bacillus clausii and Its Application in Elicitation of Plant Disease Resistance
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- 作者：Zuming Li (12)
Zhihui Bai (2) zhbai@rcees.ac.cn
Baoguo Zhang (2)
Baojv Li (3)
Bo Jin (4)
Michael Zhang (5)
Francis Lin (5)
Hongxun Zhang (2) - 关键词：Bacillus clausii – Alkaline pectin lyase – Sugar beet pulp – Mass spectrometric sequencing – Plant disease resistance
- 刊名：Applied Biochemistry and Biotechnology
- 出版年：2012
- 出版时间：August 2012
- 年：2012
- 卷：167
- 期：8
- 页码：2241-2256
- 全文大小：351.3 KB
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- ISSN：1559-0291
文摘
Alkaline pectin lyase (PNL) shows potential as a biological control agent against several plant diseases. We isolated and characterized a new Bacillus clausii strain that can produce 4,180 U/g of PNL using sugar beet pulp as a carbon source and inducer. The PNL was purified to apparent homogeneity using ultrafiltration, ammonium sulfate fractionation, DEAE Sepharose Fast Flow, and Sephadex G-75 gel filtration. The purified PNL was found to be a monomeric protein with a molecular weight of 35 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). It demonstrated optimal activity with K m of 0.87 mg/ml at pH 10.0 and 60 °C. The enzyme is stable in the pH range of 8.0–10.0 and temperature ≤40 °C. Ca2+ was found to stimulate the enzymatic activity of the PNL by up to 410 %. Mass spectrometric results gave 38 % match coverage with pectate lyase from B. clausii KSM-K16 (gi|56961845). The PNL was found to elicit disease resistance in cucumber seedlings, suggesting that it may have applications in biocontrol and sustainable agriculture.