Document Detail


Molecular cloning and characterization of trehalose biosynthesis genes from hyperthermophilic archaebacterium Metallosphaera hakonesis.
MedLine Citation:
PMID:  18051363     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The trehalose (alpha-D-glucopyranosyl-[1,1]-alpha-D-glucopyranose) biosynthesis genes MhMTS and MhMTH, encoding a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively, have been cloned from the hyperthermophilic archaebacterium Metallosphaera hakonesis. The ORF of MhMTS is 2,142 bp long, and encodes 713 amino acid residues constituting a 83.8 kDa protein. MhMTH is 1,677 bp long, and encodes 558 amino acid residues constituting a 63.7 kDa protein. The deduced amino acid sequences of MhMTS and MhMTH contain four regions highly conserved for MTSs and three for MTHs that are known to constitute substrate-binding sites of starch-hydrolyzing enzymes. Recombinant proteins obtained by expressing the MhMTS and MhMTH genes in E. coli catalyzed a sequential reaction converting maltooligosaccharides to produce trehalose. Optimum pH of the MhMTS/MhMTH enzyme reaction was around 5.0 and optimum temperature was around 70 degrees C. Trehalose-producing activity of the MhMTS/ MhMTH was notably stable, retaining 80% of the activity after preincubation of the enzyme mixture at 70 degrees C for 48 h, but was gradually abolished by incubating at above 85 degrees C. Addition of thermostable 4-alpha-glucanotransferase increased the yield of trehalose production from maltopentaose by 10%. The substrate specificity of the MhMTS/MhMTH-catalyzed reaction was extended to soluble starch, the most abundant maltodextrin in nature.
Authors:
Ju-Seok Seo; Ju Hee An; Moo-Yeol Baik; Cheon Seok Park; Jong-Joo Cheong; Tae Wha Moon; Kwan Hwa Park; Yang Do Choi; Chung Ho Kim
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microbiology and biotechnology     Volume:  17     ISSN:  1017-7825     ISO Abbreviation:  J. Microbiol. Biotechnol.     Publication Date:  2007 Jan 
Date Detail:
Created Date:  2007-12-05     Completed Date:  2008-01-22     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9431852     Medline TA:  J Microbiol Biotechnol     Country:  Korea (South)    
Other Details:
Languages:  eng     Pagination:  123-9     Citation Subset:  IM    
Affiliation:
Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921, Korea.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Cloning, Molecular
DNA Primers / genetics
DNA, Archaeal / genetics
Enzyme Stability
Escherichia coli / genetics
Genes, Archaeal*
Glucosidases / genetics,  metabolism
Glucosyltransferases / genetics,  metabolism
Glycogen Debranching Enzyme System / genetics,  metabolism
Hot Temperature
Recombinant Proteins / genetics,  metabolism
Starch / metabolism
Substrate Specificity
Sulfolobaceae / enzymology,  genetics*,  metabolism*
Thermotoga maritima / enzymology,  genetics
Trehalose / biosynthesis*,  genetics
Chemical
Reg. No./Substance:
0/DNA Primers; 0/DNA, Archaeal; 0/Glycogen Debranching Enzyme System; 0/Recombinant Proteins; 9005-25-8/Starch; 99-20-7/Trehalose; EC 2.4.1.-/Glucosyltransferases; EC 2.4.1.-/maltooligosyl trehalose synthase; EC 2.4.1.25/4 alpha-glucanotransferase; EC 3.2.1.-/Glucosidases; EC 3.2.1.141/maltooligosyl trehalose trehalohydrolase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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