Compositions and methods of use
A kind of technology of composition, enzyme composition, applied in directions such as hydrolase
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[0224] Preparation and use of replicable vectors
[0225] DNA encoding the Tfu Man1 protein or derivatives thereof (as described above) is prepared for insertion into a suitable microorganism. According to the present compositions and methods, the DNA encoding the Tfu Man1 polypeptide includes all DNA necessary to encode a protein having functional Tfu Man1 activity. Thus, embodiments of the present compositions and methods include DNA encoding a Tfu Man1 polypeptide derived from a Thermobiida spp., including Thermophilia browna.
[0226] The DNA encoding Tfu Man1 can be prepared by constructing an expression vector carrying the DNA encoding Tfu Man1. The expression vector carrying the inserted DNA segment encoding Tfu Man1 can be any vector capable of autonomous replication in a given host organism or capable of integrating into the DNA of the host, usually a plasmid, cosmid, virus particle or phage. Various vectors are publicly available. It is also envisioned that more...
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[0284] The following examples are presented to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the present compositions and methods, and are not intended to limit the scope of what the inventors believe to be their inventive compositions and methods, nor are they intended to Experiments below performance were all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, etc.), but some experimental errors and deviations should be accounted for.
example 1
[0286] Cloning of Glycosyl Hydrolase tfu Man1 from Thermobista spp.
[0287] The actinomycete Thermobifida spp. strain YX was selected as a potential source of various glycosyl hydrolases and other enzymes for industrial applications. The nucleic acid sequence of the tfu Man1 gene (NCBI accession number YP_288961) encoding Tfu Man1 was synthesized by GeneRay Biotech Co., Ltd (Shanghai, China).
[0288]TfuMan1 (NCBI accession number: YP_288961 ) is an endomannan-1,4-β-mannosidase from the actinomycete Thermobifida sp. YX. The nucleic acid sequence of the gene tfu Man1 encoding Tfu Man1 is provided herein as SEQ ID NO:1. The amino acid sequence of the protein encoded by the tfu Man1 gene is provided herein as SEQ ID NO:2. At the N-terminus, the protein is predicted to have a signal peptide of 28 amino acids in length, as determined by the Signal P 3.0 program (www.cbs.dtu / services / SignalP) programmed to the SignalP-NN system (Emanuelsson et al., Nature Protocols ("Nature La...
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