Hydroxylase gene and use thereof
AssigneesMicrobiopharm Japan Co., Ltd., National Institute of Advanced Industrial Science and Technology
InventorsYoshikazu Fujii, Tomohiro Tamura
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Abstract
A vitamin D3 hydroxylase is purified from Pseudonocardia autotrophica cell, and a primer is designed based on amino acid sequence obtained from hydroxylase. Subsequently, PCR is conducted using genomic DNA of Pseudonocardia autotrophica as a template to clone a gene for the vitamin D3 hydroxylase. By conducting a conversion reaction using a microorganism in which the vitamin D3 hydroxylase gene is expressed using a proper expression system, a hydroxide of vitamin D or the like (e.g., hydroxy vitamin D3) can be produced with high efficiency.
Claims9
An isolated polynucleotide encoding a hydroxylase selected from (a)-(c):
- (a) an isolated polynucleotide comprising nucleotides 320-1531 of SEQ ID NO:
- 1;
- and
- (b) an isolated polynucleotide that hybridizes under stringent conditions to a polynucleotide entirely complementary to nucleotides 320-1531 of SEQ ID NO:
- 1, wherein said hybridization under stringent conditions is hybridization at 65° C. in the presence of 0.7-1.0M sodium chloride;
- and
- (c) an isolated polynucleotide comprising a polynucleotide sequence having at least 90% nucleic acid sequence identity to nucleotides 320-1531 of SEQ ID NO:
- 1.
An isolated polynucleotide selected from (a) and (b):
- (a) an isolated polynucleotide comprising a polynucleotide sequence encoding the amino acid sequence of SEQ ID NO:
- 2;
- and
- (b) an isolated polynucleotide that hybridizes under stringent conditions to a polynucleotide entirely complementary to a polynucleotide encoding the amino acid sequence of SEQ ID NO:
- 2, wherein said hybridization under stringent conditions is hybridization at 65° C. in the presence of 0.7-1.0M sodium chloride, wherein the polynucleotide encodes a hydroxylase.
Dependencies are inferred from the claim text.