Botryosphaeria rhodina polypeptide
A mature polypeptide and polynucleotide technology, applied to DNA/RNA fragments, cells modified by introducing foreign genetic material, enzymes, etc.
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[0286] Compositions comprising enzyme polypeptides and methods for their preparation
[0287] The invention provides compositions comprising a polypeptide of the invention and an excipient and methods of preparing such compositions comprising admixing a polypeptide of the invention and an excipient. In a specific embodiment, the polypeptide of the invention is the main (polypeptide) component of the composition, eg a one-component composition. Here, an excipient should be understood as any auxiliary agent or compound used to formulate the composition, including solvents, carriers, stabilizers and the like.
[0288] The composition may also contain one or more other enzymes such as aminopeptidase, amylase, carbohydrase, carboxypeptidase, catalase, cellulase, chitinase, cutinase, cyclodextrin sugar Base transferase, deoxyribonuclease, esterase, α-galactosidase, β-galactosidase, glucoamylase, α-glucosidase, β-glucosidase, haloperoxidase ), invertase, laccase, lipase, mannosidas...
Embodiment 1
[0341] Example 1 Identification of functional polypeptides secreted by Botryosphaeria rhodina CBS 274.96
[0342] Enzyme fingerprint analysis of culture medium
[0343] Enzyme activity profiles were obtained by assaying the culture broths with various enzyme assays. 96-well microtiter (MT) plates with substrate were prepared and stored at 10°C until use. Two different pH variations were prepared: pH3 and pH7. The following substrates were used: 0.05% AZCL (Mazurine staining and cross-linking substrate, Megazyme) - amylose, arabinan, β-glucan (barley), casein, collagen, curdlan, dextran Sugars, galactan (potato), galactomannan (carob) (carob), He-cellulose, pullulan (pullulan), xylan (oat) and xyloglucan (AZCL-casein Egg whites cannot be used at pH 3, so no AZCL-casein was added to these plates).
[0344] Preparation of pH3 substrate:
[0345] 0.1 g of each AZCL substrate was dissolved in 100 ml of 0.2M succinic acid pH 3 + 10 microliters of Triton X-100 (0.01%) to give a ...
Embodiment 2
[0402] Example 2 Determination of Enzyme Activity by Homology
[0403] The function of a gene or encoded polypeptide can be predicted by comparison with genes or polypeptides of known function. Analyzed a DNA Group and B 多肽 Similarity between group sequences and sequences from public and internal databases to determine a DNA Group and B 多肽 Functionality of group sequences. Sequence comparisons were performed with the program BLASTX2.0a19MP-WashU [1998-7-14]. by adding A DNA Group and B 多肽 Careful manual alignment of the group sequence with its closest sequence of known function provides the possibility to predict the function of these genes and encoded polypeptides. Even in cases where overall amino acid identity is below 40%, making reliable predictions may be difficult, by careful analysis and interpretation of amino acid residues in important regions of the catalytic site and / or polypeptide sequence, A DNA Group and B 多肽 Functions of group sequences are also possib...
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