Cloning and characterization of glucokinase from a methylotrophic yeast Hansenula polymorpha : different effects on glucose repression in H . polymorpha and Saccharomyces cerevisiae
| Autor: | Aiki Järviste, Helen Karp, Pille Kotka, Tiina Alamäe, Silja Laht |
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| Rok vydání: | 2002 |
| Předmět: |
Molecular Sequence Data
Saccharomyces cerevisiae Mutant Mannose Pichia Substrate Specificity chemistry.chemical_compound Transformation Genetic Glucokinase Genetics Amino Acid Sequence Cloning Molecular Phosphorylation DNA Fungal Phylogeny Genomic Library Hexokinase Base Sequence Sequence Homology Amino Acid biology Sequence Analysis DNA General Medicine biology.organism_classification Molecular biology Complementation Glucose Invertase chemistry Biochemistry Mutation Maltase Sequence Alignment Plasmids |
| Zdroj: | Gene. 296:195-203 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/s0378-1119(02)00859-4 |
| Popis: | Glucokinase gene (HPGLK1) was cloned from a methylotrophic yeast Hansenula polymorpha by complementation of glucose-phosphorylation deficiency in a H. polymorpha double kinase-negative mutant A31-10 by a genomic library. An open reading frame of 1416 nt encoding a 471-amino-acid protein with calculated molecular weight 51.6 kDa was characterized in the genomic insert of the plasmid pH3. The protein sequence deduced from HPGLK1 exhibited 55 and 46% identity with glucokinases from Saccharomyces cerevisiae and Aspergillus niger, respectively. The enzyme phosphorylated glucose, mannose and 2-deoxyglucose, but not fructose. Transformation of HPGLK1 into A31-10 restored glucose repression of alcohol oxidase and catalase in the mutant. Transformation of HPGLK1 into S. cerevisiae triple kinase-negative mutant DFY632 showed that H. polymorpha glucokinase cannot transmit the glucose repression signal in S. cerevsiae: synthesis of invertase and maltase in respective transformants was insensitive to glucose repression similarly to S. cerevisiae DFY568 possessing only glucokinase. |
| Databáze: | OpenAIRE |
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