Hyaluronan synthase genes and expression thereof in Bacillus hosts
OwnerPaul DeAngelis, Paul Weigel +1inventors
InventorsPaul DeAngelis, Paul Weigel, Kshama Kumari
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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).
Claims20
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.
A method for producing hyaluronic acid, comprising the steps of:
- 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.
Dependencies are inferred from the claim text.