Genetic Engineering of Stenotrophomonas Strain YC-1 To Possess a Broader Substrate Range for Organophosphates

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• 作者：Chao Yang ; Cunjiang Song ; Ashok Mulchandani ; Chuanling Qiao
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2010
• 出版时间：June 9, 2010
• 年：2010
• 卷：58
• 期：11
• 页码：6762-6766
• 全文大小：722K
• 年卷期：v.58,no.11(June 9, 2010)
• ISSN：1520-5118

文摘

In this work, Stenotrophomonas sp. strain YC-1, a native soil bacterium that produces methyl parathion hydrolase (MPH), was genetically engineered to possess a broader substrate range for organophosphates (OPs). A surface anchor system derived from the truncated ice nucleation protein (INPNC) from Pseudomonas syringae was used to target organophosphorus hydrolase (OPH) onto the cell surface of strain YC-1, reducing the potential substrate uptake limitation. The surface localization of INPNC−OPH was verified by cell fractionation, Western blot, proteinase accessibility, and immunofluorescence microscopy. No growth inhibition was observed for the engineered strain, and suspended cultures retained almost 100% activity over a period of 2 weeks. Concomitant expression of OPH in strain YC-1 resulted in a recombinant strain capable of simultaneously degrading diethyl and dimethyl OPs. A mixture of six OP pesticides (0.2 mM each) could be degraded completely within 5 h. The broader substrate specificity in combination with the rapid degradation rate makes this engineered strain a promising candidate for in situ remediation of OP-contaminated sites.