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Improved bacillus promoter

IP.com Disclosure Number: IPCOM000241578D
Publication Date: 2015-May-13

Publishing Venue

The IP.com Prior Art Database

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RESEARCH DISCLOSURE: IMPROVED BACILLUS PROMOTER

SUMMARY

     This research disclosure provides a prokaryotic host cell for producing a polypeptide of interest, said host cell comprising a polynucleotide construct which comprises:

(a) a quadruple promoter comprising four or more promoter sequences, wherein the quadruple promoter is operably linked to a polynucleotide encoding the polypeptide of interest, and

(b) an mRNA processing/stabilizing sequence located downstream of the quadruple promoter and upstream of the polypeptide-encoding polynucleotide,

wherein at least one of the promoters of the quadruple promoter is selected from the group

consisting of an E. coli lac operon promoter, a Streptomyces coelicolor agarase gene (dagA) promoter, a Bacillus lentus alkaline protease gene (aprH) promoter, a Bacillus licheniformis alkaline protease gene promoter, a Bacillus subtilis levansucrase gene (sacB) promoter, a Bacillus subtilis alpha-amylase gene (amyE) promoter, a Bacillus licheniformis alpha-amylase gene (amyL) promoter, a Bacillus stearothermophilus maltogenic amylase gene (amyM)

promoter, a Bacillus amyloliquefaciens alpha-amylase gene (amyQ) promoter, a Bacillus licheniformis penicillinase gene (penP) promoter, a Bacillus subtilis xylA gene promoter, a Bacillus subtilis xylB gene promoter, a prokaryotic beta-lactamase gene promoter, a spo1 bacterial phage promoter, a bacterial tac promoter and a "consensus" promoter having the sequence TTGACA for the "-35" region and TATAAT for the "-10" region.

     The host cell is preferably a Bacillus host cell; preferably the Bacillus host cell is selected from the group consisting of Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis and Bacillus thuringiensis.

     The polypeptide of interest is preferably heterologous or homologous to the host cell. Ideally, the polypeptide of interest is secreted; preferably the polypeptide of interest is an enzyme; more preferably the enzyme is an oxidoreductase, transferase, hydrolase, lyase, isomerase, or ligase; and most preferably the enzyme is an aminopeptidase, amylase, carbohydrase, carboxypeptidase, catalase, cellulase, chitinase, cutinase, cyclodextrin

glycosyltransferase, deoxyribonuclease, esterase, alpha-galactosidase, beta-galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, invertase, laccase, lipase, mannosidase, mutanase, oxidase, a pectinolytic enzyme, peroxidase, phytase, polyphenoloxidase, protease, ribonuclease, transglutaminase, or xylanase.

It may also be that the polypeptide of interest is intracellular; preferably the polypeptide

of interest is involved in the synthesis or degradation of a compound; more preferably the polypeptide of interest is a hyaluronic acid synthase.

     The quadruple promoter...