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Methods and nucleic acid sequences for the expression of the cellulose synthase operon
| Details |
Inventors: Ben-Bassat, Arie; Calhoon, Roger D.; Fear, Anna L.; Gelfand, David H.; Meade, James H.; Tal, Rony; Wong, Hing; Benziman, Moshe;
Assignee: Cetus Corporation (Emeryville, CA)
Primary Examiner: Wax; Robert A.
Assistant Examiner: Moore; William W.
Attorney, Agent or Firm: Halluin; Albert P.
Nucleic acid sequences encoding the bacterial cellulose synthase operon derived from Acetobacter are disclosed. Methods for isolating the genes, vectors containing the genes, and transformed hosts useful for the expression of recombinant bacterial cellulose synthase or production of cellulose are also described. |
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DETAILED DESCRIPTION The present invention provides an operon associated with the biosynthesis of cellulose, polynucleotides encoding one or more closely linked genes that code for proteins of bacterial cellulose synthase, expression vectors suitable for production of cellulose synthase recombinant host cells transformed with these vectors, methods for producing bacterial cellulose synthase, methods for regulating the production of cellulose and methods to increase the production of cellulose in a recombinant microorganism.
More particularly the invention provides an isolated native, cloned recombinant or synthetic polynucleotide encoding the bacterial cellulose synthase operon characterized by the polycistronic nucleotide sequence shown in FIG.
1 (SEQ ID No: 1).
The cellulose synthase operon is approximately 9217 basepairs (bp) in length and comprises four genes, designated herein as "A" (SEQ ID No: 3), "B" (SEQ ID No: 4), "C" (SEQ ID No: 5), and "D" (SEQ ID No: 6).
The invention further provides a process for expressing cellulose synthase in a host cell comprising transforming the host cell with a recombinant DNA expression vector comprising one or more of the genes associated with the bacterial cellulose synthase operon, which gene(s) is operably linked to a control sequence for expression of bacterial cellulose synthase, and culturing the transformed host cell under conditions suitable for expression of cellulose synthase.
The expression vector constructions of the present invention can either replicate independently or may be designed so as to introduce a heterologous promoter into the Acetobacter chromosome, thereby replacing the native cellulose synthase operon promoter.
Yet another aspect of the invention provides a method for increasing cellulose production in a recombinant microorganism, which method comprises transforming a suitable microorganism with a vector comprising at least one gene derived from the cellulose synthase operon, and culturing the transformed microorganism under conditions suitable for production of cellulose
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Molecular Biology 
