Document Detail


Expression and characterization of sweet potato invertase in Pichia pastoris.
MedLine Citation:
PMID:  12590504     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An invertase cDNA (Ibbetafruct1) was cloned from sweet potato leaves and characterized. The deduced amino acid sequence of the Ibbetafruct1-encoded protein was closely related to vacuolar invertases and included the WECVD catalytic domain characteristic of them. An expression plasmid containing the coding region of Ibbetafruct1 under the control of the alcohol oxidase promoter was used to transform the methylotrophic yeast Pichia pastoris. The biochemical properties for the expressed recombinant enzyme, which was determined to be the acid beta-fructofuranosidase with an acidic pI value (5.1), were similar to those of vacuolar invertases purified from sweet potato. Periodic acid/Schiff staining and Con A-Sepharose gel-binding experiments revealed the recombinant invertase to be a glycoprotein containing glucose and/or mannose residues. Furthermore, the carbohydrate moiety appears to be a key determinant of the enzyme's sucrose hydrolysis activity, substrate affinity, and thermal stability.
Authors:
Wen-Chin Huang; Ai-Yu Wang; Li-Ting Wang; Hsien-Yi Sung
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of agricultural and food chemistry     Volume:  51     ISSN:  0021-8561     ISO Abbreviation:  J. Agric. Food Chem.     Publication Date:  2003 Feb 
Date Detail:
Created Date:  2003-02-19     Completed Date:  2003-04-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0374755     Medline TA:  J Agric Food Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1494-9     Citation Subset:  IM    
Affiliation:
Department of Agricultural Chemistry, National Taiwan University, No. 1, Section 4, Roosevelt Road, Taipei, Taiwan.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Cloning, Molecular
DNA, Complementary / chemistry,  genetics
DNA, Plant / genetics
Gene Expression*
Glycoside Hydrolases / chemistry,  genetics*,  metabolism
Glycosylation
Ipomoea batatas / enzymology*,  genetics*
Molecular Sequence Data
Pichia / enzymology,  genetics*
Plant Leaves / enzymology
Recombinant Proteins / metabolism
Sequence Alignment
Transfection
beta-Fructofuranosidase
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/DNA, Plant; 0/Recombinant Proteins; EC 3.2.1.-/Glycoside Hydrolases; EC 3.2.1.26/beta-Fructofuranosidase

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


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