Construction and application of curvularia lunata steroid C14 alpha-hydroxylase CYP5103B5 mutant

The invention provides curvularia lunata steroid C14 alpha-hydroxylase mutants Y10 (S129A/T130A/D131G/T132L/V133I/R134K/S135K) and D131G, and the coding nucleotide sequences of the curvularia lunata steroid C14 alpha-hydroxylase mutants Y10 (S129A/T130A/D131G/T132L/V133I/R134K/S135K) and D131G are s...

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Hauptverfasser:	WANG SHUO, MAO SHUHONG, LIU XIAOGUANG, QIAO JIANGYU, LU FUPING
Format:	Patent
Sprache:	chi ; eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROCESSES USING MICROORGANISMS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Zusammenfassung:	The invention provides curvularia lunata steroid C14 alpha-hydroxylase mutants Y10 (S129A/T130A/D131G/T132L/V133I/R134K/S135K) and D131G, and the coding nucleotide sequences of the curvularia lunata steroid C14 alpha-hydroxylase mutants Y10 (S129A/T130A/D131G/T132L/V133I/R134K/S135K) and D131G are shown as SEQ ID NO: 4 and SEQ ID NO: 6. Compared with C14 alpha-hydroxylase CYP5103B5, the mutant Y10 (S129A/T130A/D131G/T132L/V133I/R134K/S135K) and D131G have the advantages that the specificity of conversion of steroid substrate progesterone is obviously improved, and by-products are obviously reduced. The invention further provides an expression vector for heterologous overexpression of the CYP5103B5 mutant and a steroid transformation process of the expression vector, and valuable materials (genes and strains) and basic data support are provided for research and development of an efficient progesterone C14 alpha-hydroxylation production process. 本发明专利提供了新月弯孢霉甾体C14α-羟化酶突变体Y10(S129A/T130A/D131G/T132L/V133I/R134K/