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Author Li, Jianfang ♦ Tang, Cunduo ♦ Shi, Hongling ♦ Wu, Minchen
Source World Health Organization (WHO)-Global Index Medicus
Content type Text
Publisher Elsevier
File Format HTM / HTML
Language English
Difficulty Level Medium
Subject Domain (in DDC) Natural sciences & mathematics ♦ Chemistry & allied sciences ♦ Life sciences; biology ♦ Physiology & related subjects ♦ Biochemistry ♦ Genetics and evolution ♦ Natural history of organisms ♦ Technology ♦ Medicine & health ♦ Human physiology ♦ Pharmacology and therapeutics ♦ Diseases ♦ Chemical engineering ♦ Manufacture for specific uses ♦ Precision instruments & other devices
Subject Domain (in MeSH) Eukaryota ♦ Organisms ♦ Enzymes and Coenzymes ♦ Amino Acids, Peptides, and Proteins ♦ Chemicals and Drugs ♦ Investigative Techniques ♦ Analytical, Diagnostic and Therapeutic Techniques and Equipment ♦ Chemical Phenomena ♦ Genetic Phenomena ♦ Biological Sciences
Subject Keyword Discipline Biomedical Engineering ♦ Discipline Microbiology ♦ Cellulase ♦ Metabolism ♦ Fungal Proteins ♦ Volvariella ♦ Enzymology ♦ Genetics ♦ Cloning, Molecular ♦ Electrophoresis, Polyacrylamide Gel ♦ Electroporation ♦ Enzyme Stability ♦ Gene Expression ♦ Glycosylation ♦ Hydrogen-ion Concentration ♦ Pichia ♦ Sequence Homology, Amino Acid ♦ Journal Article
Abstract A cDNA fragment encoding a mature neutral endoglucanase with 366 amino acids was cloned from Volvariella volvacea WX32. Online analysis of amino acid sequence homology showed that the endoglucanase, designated as NCel5A, belongs to glycoside hydrolase family 5. The recombinant plasmid, pPIC9K-ncel5A, was transformed into Pichia pastoris GS115 by electroporation. Screening of multiple copies of the gene ncel5A in transformants was performed on YPD plates containing geneticin G418. One transformant expressing the highest recombinant NCel5A (rNCel5A) activity, numbered as GSNCel4-3, was chosen for optimizing expression conditions. In optimized conditions, the expressed rNCel5A activity was up to 4612 U/ml. SDS-PAGE and enzyme activity assays demonstrated that the rNCel5A, a glycosylated protein with an M.W. of about 42 kDa, was extracellularly expressed in P. pastoris. The rNCel5A showed the highest activity at pH 7.5 and 55°C and was stable at a broad pH range of 6.0-9.0 and at a temperature of 55°C or below.
Description Author Affiliation: Li J ( School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, PR China.)
ISSN 13891723
Educational Role Student ♦ Teacher
Age Range above 22 year
Educational Use Reading ♦ Research ♦ Self Learning
Interactivity Type Expositive
Education Level UG and PG
Learning Resource Type Article
Publisher Date 2011-05-01
Publisher Place Japan
e-ISSN 13474421
Journal Journal of Bioscience and Bioengineering
Volume Number 111
Issue Number 5

Source: WHO-Global Index Medicus