Wilmar (shanghai) Biotechnology Research & Development Center Co., Ltd.

Qiwen Niu

Wei Wu

Yaoji Xuan

Haisheng Cao

Zhengjun Xu

Xiaojun Dai

Sha Liu

Meifeng Zhou

Yueyi Bao

Sitian Gu

Provided is amutant strain for expressing an exogenous gene. Specifically, provided is amutant strain comprising, with respect tomutant strain GS115 or CICC32806, one or more of the following six mutations: BQ9382_C1-2260, EKK deletions at positions 308-310, a hypothetical protein; BQ9382_C1-3800, E129K, 60S ribosomal subunit assembly/exported protein LOC1; BQ9382_C1-5700, 1312M, mitochondrial external NADH dehydrogenase, type II NAD(P)H: quinone oxidoreductase; BQ9382_C2-3950, Q145X, an essential protein having a binding partner Psr1p and used for completely activating a general stress response; BQ9382_C3-2220, E188K, a hypothetical protein; and BQ9382_C3-4370, W196X, orotidine 5\'-phosphate decarboxylase. The providedmutant strain is an effective commonly employed host for exogenous expression, and can efficiently express different proteins, especially phospholipase and lipase.

mutant strain for expressing exogenous gene

Provided is a polypeptide having phospholipase C activity. The polypeptide has: 1) an amino acid sequence represented by SEQ ID No: 2, comprising amino acid substitutions occurred at one or more positions, wherein the one or more positions are selected from positions 6, 8, 10, 104, and 205 in the amino acid sequence represented by SEQ ID No: 2 or any combination thereof; or 2) having at least 80% sequence identity with 1), and at least one of positions 6, 8, 10, 104 and 205 being different from positions 6, 8, 10, 104, and 205 in the amino acid sequence represented by SEQ ID No: 2. Provided are a nucleic acid molecule for encoding the polypeptide, a vector comprising the nucleic acid molecule, and a cell comprising the nucleic acid molecule or the vector. Provided are uses of the polypeptide, the nucleic acid molecule, the vector, and the cell.

Polypeptide having phospholipase C activity and use thereof

Provided in the present invention is a phospholipase C mutant with high enzyme activity, a polypeptide with the sequence as shown in SEQ ID NO:7, or a polypeptide derived from the phospholipase C formed by performing substitution, deletion or addition of one or a plurality of amino acids on the polypeptide of SEQ ID NO:7 while retaining the phospholipase C activity provided by SEQ ID NO:7. The phospholipase C mutant of the present invention can improve degumming efficiency and increase the yield of diacylglycerol (DAG) during degumming.

Phospholipase C mutant with high enzyme activity

According to the present disclosure, on the basis of all existing mutations, the tenth glycine of a BC-PC-PLC is mutated into aspartic acid, a specific enzyme activity thereof is 83% higher than that of a sequence before the mutation, and protein expression and degumming activity of unit enzyme activity do not change, so as to further reduce manufacturing costs.

Phospholipase C and encoding gene thereof

Provided is a mutant of the wild type phosphatidylcholine-specific phospholipase C of. The mutations involved comprise the amino acid residue at position 63 being mutated from asparagine to aspartic acid, the amino acid residue at position 131 being mutated from asparagine to serine, and the amino acid residue at position 134 being mutated from asparagine to aspartic acid, and may comprise the amino acid residue at position 56 being mutated from tyrosine to alanine, lysine, asparagine, glutamine, histidine or tryptophan, and further, may also comprise the amino acid residue at position 106 being mutated from methionine to valine. Also provided are a polynucleotide molecule encoding the mutant, a nucleic acid construct and a host cell comprising the polynucleotide molecule, a composition comprising the mutant, and the use of the mutant, the polynucleotide molecule, the nucleic acid construct and the host cell.

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Shanghai, China

Qiwen Niu