| Molecular cloning and functional analysis of (R)-3-hydroxyacyl-acyl carrier protein:coenzyme A transacylase from Pseudomonas mendocina LZ. | |
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MedLine Citation:
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PMID: 16213672 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An inactive (R)-3-hydroxyacyl-acyl carrier protein:coenzyme A transacylase (PhaG(Pm)) was cloned from a newly isolated Proteobacteria Pseudomonas mendocina LZ. It is the first characterized native inactive PhaG protein. Sequence analysis indicated that there were only two sites where the amino acid sequence differed between this inactive protein and the functional PhaG(Pp) from P. putida. The differences were located at position 78 and in the region 109-113 in the amino acid sequence. Mutagenesis was carried out to investigate these two sites. A recombinant strain harboring a S78C PhaG(Pp) mutant accumulated polyhydroxyalkanoates (PHA) at 11.9% of the cellular dry weight, as compared to the 21.6% PHA produced by the recombinant harboring the wild-type PhaG(Pp). On the other hand, the changes in the amino acid region 109-113 of PhaG(Pp) to its corresponding region of PhaG(Pm) resulted in negligible PHA accumulation. This demonstrated that region 109-113 in PhaG is relatively important for transacylase activity, while position 78 just plays a supporting role for the enzyme. Furthermore, 3-D structural models of PhaG(Pp) and PhaG(Pm) developed by computational prediction revealed that the variation in amino acids at 109-113 leads to the destruction of the PhaG catalytic center, resulting in the loss of enzyme activity. |
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Authors:
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Leo Zhong Zheng; Zhi Li; Hong-Lei Tian; Ming Li; Guo-Qiang Chen |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2005-09-22 |
Journal Detail:
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Title: FEMS microbiology letters Volume: 252 ISSN: 0378-1097 ISO Abbreviation: FEMS Microbiol. Lett. Publication Date: 2005 Nov |
Date Detail:
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Created Date: 2005-10-31 Completed Date: 2006-03-16 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7705721 Medline TA: FEMS Microbiol Lett Country: Netherlands |
Other Details:
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Languages: eng Pagination: 299-307 Citation Subset: IM |
Affiliation:
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MOE laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, China. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/AY338498 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acyltransferases
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chemistry*,
genetics* Amino Acid Sequence Amino Acid Substitution Bacterial Proteins / genetics Catalytic Domain Cloning, Molecular* DNA, Bacterial / chemistry, genetics Genes, Bacterial Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Mutation, Missense Protein Conformation Pseudomonas mendocina / enzymology*, genetics Sequence Analysis, DNA Sequence Homology, Amino Acid |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/DNA, Bacterial; EC 2.3.-/Acyltransferases; EC 2.3.1.-/3-hydroxyacyl-CoA-acyl carrier protein transferase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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