Hyaluronan synthase genes and expression thereof in Bacillus hosts
First Claim
1. A recombinant host cell, wherein the recombinant host cell is a Bacillus cell comprising a recombinant vector comprising a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA, and wherein the coding region is under control of a promoter, and wherein the coding region encoding enzymatically active hyaluronan synthase is selected from the group consisting of:
- (A) a coding region encoding enzymatically active hyaluronan synthase that is at least 80% identical to SEQ ID NO;
10; and
(B) a coding region that is capable of hybridizing to a complementary sequence of SEQ ID NO;
9 under hybridization conditions of 1.2-1.8×
High Phosphate Buffer (HPB) at 40-50°
C.
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Abstract
The present invention relates to a recombinant Bacillus host cell containing a recombinant vector including a nucleic acid segment having a coding region segment encoding enzymatically active hyaluronan synthase (HAS). The recombinant Bacillus host cell is utilized in a method for producing hyaluronic acid (HA).
36 Citations
20 Claims
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1. A recombinant host cell, wherein the recombinant host cell is a Bacillus cell comprising a recombinant vector comprising a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA, and wherein the coding region is under control of a promoter, and wherein the coding region encoding enzymatically active hyaluronan synthase is selected from the group consisting of:
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(A) a coding region encoding enzymatically active hyaluronan synthase that is at least 80% identical to SEQ ID NO;
10; and
(B) a coding region that is capable of hybridizing to a complementary sequence of SEQ ID NO;
9 under hybridization conditions of 1.2-1.8×
High Phosphate Buffer (HPB) at 40-50°
C. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for producing hyaluronic acid, comprising the steps of:
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introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus host wherein the coding region is under control of a promoter, and wherein the enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA, and wherein the coding region encoding enzymatically active hyaluronan synthase is selected from the group consisting of;
(A) a coding region encoding enzymatically active hyaluronan synthase that is at least 80% identical to SEQ ID NO;
10; and
(B) a coding region that is capable of hybridizing to a complementary sequence of SEQ ID NO;
9 under hybridization conditions of 1.2-1.8×
High Phosphate Buffer (HPB) at 40-50°
C.;
growing the host organism in a medium to secrete hyaluronic acid; and
recovering the secreted hyaluronic acid. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification