摘要:
The present invention provides a method for preparing a variant lipid acyltransferase enzyme by expressing a nucleotide sequence encoding a lipid acyltransferase which may comprise at least one modification at a position(s) which corresponds in the encoded amino acid sequence to an amino acid(s) located in a) the canyon region of the enzyme (i.e. preferably amino acid residues 31, 27, 85, 86, 119, and 120); and/or b) insertion site 1 (i.e. amino acid residues 22-36) and/or c) insertion site 2 (i.e. amino acid residues 74-88), wherein the canyon region, insertion site 1 and/or insertion site 2 are defined as that region which when aligned based on primary or tertiary structure corresponds to the canyon region, insertion site 1 or insertion site 2 (or the corresponding amino acid residues taught above) of the enzyme shown herein as SEQ ID No. 16 or 6 in a host organism.
摘要翻译:本发明提供了一种通过表达编码脂质酰基转移酶的核苷酸序列来制备变体脂质酰基转移酶的方法,所述脂质酰基转移酶可以在编码的氨基酸序列中相应于氨基酸的位置包含至少一个修饰, 位于a)谷的峡谷区域(即优选氨基酸残基31,27,85,86,119和120); 和/或b)插入位点1(即氨基酸残基22-36)和/或c)插入位点2(即氨基酸残基74-88),其中峡谷区域,插入位点1和/或插入位点2是 定义为当基于一级或三级结构对准的区域对应于本文所示的酶的SEQ ID No.16或6的峡谷区域,插入位点1或插入位点2(或上文所述的相应氨基酸残基) 宿主生物。
摘要:
The present invention provides a method for preparing a variant lipid acyltransferase enzyme by expressing a nucleotide sequence encoding a lipid acyltransferase which may comprise at least one modification at a position(s) which corresponds in the encoded amino acid sequence to an amino acid(s) located in a) the canyon region of the enzyme (i.e. preferably amino acid residues 31, 27, 85, 86, 119, and 120); and/or b) insertion site 1 (i.e. amino acid residues 22-36) and/or c) insertion site 2 (i.e. amino acid residues 74-88), wherein the canyon region, insertion site 1 and/or insertion site 2 are defined as that region which when aligned based on primary or tertiary structure corresponds to the canyon region, insertion site 1 or insertion site 2 (or the corresponding amino acid residues taught above) of the enzyme shown herein as SEQ ID No. 16 or 6 in a host organism.
摘要翻译:本发明提供了一种通过表达编码脂质酰基转移酶的核苷酸序列来制备变体脂质酰基转移酶的方法,所述脂质酰基转移酶可以在编码的氨基酸序列中相应于氨基酸的位置包含至少一个修饰, 位于a)谷的峡谷区域(即优选氨基酸残基31,27,85,86,119和120); 和/或b)插入位点1(即氨基酸残基22-36)和/或c)插入位点2(即氨基酸残基74-88),其中峡谷区域,插入位点1和/或插入位点2是 定义为当基于一级或三级结构对准的区域对应于本文所示的酶的SEQ ID No.16或6的峡谷区域,插入位点1或插入位点2(或上文所述的相应氨基酸残基) 宿主生物。
摘要:
A fungal wild-type lipolytic enzyme having a higher ratio of activity on polar lipids compared with triglycerides, wherein the enzyme preferably has a phospholipid:triglyceride activity ratio of at least 4. Preferably, the lipolytic enzyme according to the present invention has a glycolipid:triglyceride hydrolyzing activity ratio of at least 1.5. In one embodiment, the fungal lipolytic enzyme according to the present invention comprises an amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID No. 2 or SEQ ID No. 4 or SEQ ID No. 6 or an amino acid sequence which has at least 90% identity thereto. The present invention further encompasses a nucleic acid encoding a fungal lipolytic enzyme, which nucleic acid is selected from the group consisting of: (a) a nucleic acid comprising a nucleotide shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7; (b) a nucleic acid which is related to the nucleotide sequence of SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7 by the degeneration of the genetic code; and (c) nucleic acid comprising a nucleotide sequence which has at least 90% identity with the nucleotide sequence shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7.
摘要翻译:与甘油三酯相比,极性脂质活性比例更高的真菌野生型脂肪分解酶,其中所述酶优选具有至少为4的磷脂:甘油三酯活性比。优选地,根据本发明的脂肪分解酶具有糖脂: 甘油三酯水解活性比至少为1.5。 在一个实施方案中,根据本发明的真菌脂肪分解酶包含如SEQ ID NO:1或SEQ ID No.2或SEQ ID No.4或SEQ ID No.6所示的氨基酸序列或氨基酸序列, 与其具有至少90%的同一性。 本发明还包括编码真菌脂肪分解酶的核酸,该核酸选自:(a)包含SEQ ID No.3,SEQ ID No.5或SEQ ID No.5所示核苷酸的核酸 第7号 (b)通过遗传密码的退化与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7的核苷酸序列有关的核酸; 和(c)核酸,其包含与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7所示的核苷酸序列具有至少90%同一性的核苷酸序列。
摘要:
A fungal wild-type lipolytic enzyme having a higher ratio of activity on polar lipids compared with triglycerides, wherein the enzyme preferably has a phospholipid:triglyceride activity ratio of at least 4. Preferably, the lipolytic enzyme according to the present invention has a glycolipid:triglyceride hydrolyzing activity ratio of at least 1.5. In one embodiment, the fungal lipolytic enzyme according to the present invention comprises an amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID No. 2 or SEQ ID No. 4 or SEQ ID No. 6 or an amino acid sequence which has at least 90% identity thereto. The present invention further encompasses a nucleic acid encoding a fungal lipolytic enzyme, which nucleic acid is selected from the group consisting of: (a) a nucleic acid comprising a nucleotide shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7; (b) a nucleic acid which is related to the nucleotide sequence of SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7 by the degeneration of the genetic code; and (c) nucleic acid comprising a nucleotide sequence which has at least 90% identity with the nucleotide sequence shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7.
摘要翻译:与甘油三酯相比,极性脂质活性比例更高的真菌野生型脂肪分解酶,其中所述酶优选具有至少为4的磷脂:甘油三酯活性比。优选地,根据本发明的脂肪分解酶具有糖脂: 甘油三酯水解活性比至少为1.5。 在一个实施方案中,根据本发明的真菌脂肪分解酶包含如SEQ ID NO:1或SEQ ID No.2或SEQ ID No.4或SEQ ID No.6所示的氨基酸序列或氨基酸序列, 与其具有至少90%的同一性。 本发明还包括编码真菌脂肪分解酶的核酸,该核酸选自:(a)包含SEQ ID No.3,SEQ ID No.5或SEQ ID No.5所示核苷酸的核酸 第7号 (b)通过遗传密码的退化与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7的核苷酸序列有关的核酸; 和(c)核酸,其包含与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7所示的核苷酸序列具有至少90%同一性的核苷酸序列。
摘要:
A fungal wild-type lipolytic enzyme having a higher ratio of activity on polar lipids compared with triglycerides, wherein the enzyme preferably has a phospholipid:triglyceride activity ratio of at least 4. Preferably, the lipolytic enzyme according to the present invention has a glycolipid:triglyceride hydrolyzing activity ratio of at least 1.5. In one embodiment, the fungal lipolytic enzyme according to the present invention comprises an amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID No. 2 or SEQ ID No. 4 or SEQ ID No. 6 or an amino acid sequence which has at least 90% identity thereto. The present invention further encompasses a nucleic acid encoding a fungal lipolytic enzyme, which nucleic acid is selected from the group consisting of: (a) a nucleic acid comprising a nucleotide shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7; (b) a nucleic acid which is related to the nucleotide sequence of SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7 by the degeneration of the genetic code; and (c) nucleic acid comprising a nucleotide sequence which has at least 90% identity with the nucleotide sequence shown in SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7.
摘要翻译:与甘油三酯相比,极性脂质活性比例更高的真菌野生型脂肪分解酶,其中所述酶优选具有至少为4的磷脂:甘油三酯活性比。优选地,根据本发明的脂肪分解酶具有糖脂: 甘油三酯水解活性比至少为1.5。 在一个实施方案中,根据本发明的真菌脂肪分解酶包含如SEQ ID NO:1或SEQ ID No.2或SEQ ID No.4或SEQ ID No.6所示的氨基酸序列或氨基酸序列, 与其具有至少90%的同一性。 本发明还包括编码真菌脂肪分解酶的核酸,该核酸选自:(a)包含SEQ ID No.3,SEQ ID No.5或SEQ ID No.5所示核苷酸的核酸 第7号 (b)通过遗传密码的退化与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7的核苷酸序列有关的核酸; 和(c)核酸,其包含与SEQ ID No.3,SEQ ID No.5或SEQ ID No.7所示的核苷酸序列具有至少90%同一性的核苷酸序列。
摘要:
The present application provides a method of reducing and/or removing diglyceride from an edible oil, comprising admixing an edible oil with an acyl acceptor substrate and a diglyceride:glycerol acyltransferase, wherein the diglyceride:glycerol acyltransferase is characterized as an enzyme which in an edible oil is capable of transferring an acyl group from a diglyceride to glycerol. The diglyceride:glycerol acyltransferase can comprise the amino acid sequence motif GDSX. The present invention also relates to the use of a diglyceride:glycerol acyltransferase in the manufacture of an edible oil, for reducing and/or removing diglyceride from said edible oil and to the use of said enzyme in the manufacture of a foodstuff comprising an edible oil for improving the crystallization properties of said foodstuff.
摘要:
A method of producing one or more of a carbohydrate ester, a protein ester, a protein subunit ester or a hydroxyl acid ester, which method comprises admixing an acyl donor, an acyl acceptor and water to produce a high water environment comprising 5-98% water, wherein said acyl donor is a lipid substrate selected from one or more of the group consisting of a phospholipid, a lysophospholipid, a triacylglyceride, a diglyceride, a glycolipid or a lysoglycolipid and said acyl acceptor is selected from one ore more of the group consisting of a carbohydrate, a protein, a protein subunit, or a hydroxyl acid; and contacting the admixture with a lipid acyltransferase, such that said lipid acyl transferase catalyses one or both of the following reactions: alcoholysis or transesterification.
摘要:
Provided is a method of producing one or more of a carbohydrate ester, a protein ester, a protein subunit ester or a hydroxyl acid ester. The method comprises admixing an acyl donor, an acyl acceptor and water to produce a high water environment comprising 5-98% water. Preferably, the acyl donor is a lipid substrate selected from one or more of the group consisting of a phospholipid, a lysophospholipid, a triacylglyceride, a diglyceride, a glycolipid or a lysoglycolipid. Preferably, the acyl acceptor is selected from one ore more of the group consisting of a carbohydrate, a protein, a protein subunit, or a hydroxyl acid. The method further comprises contacting the admixture with a lipid acyltransferase, such that said lipid acyl transferase catalyses alcoholysis and/or transesterification.
摘要:
A method of producing one or more of a carbohydrate ester, a protein ester, a protein subunit ester or a hydroxyl acid ester, which method comprises admixing an acyl donor, an acyl acceptor and water to produce a high water environment comprising 5-98% water, wherein said acyl donor is a lipid substrate selected from one or more of the group consisting of a phospholipid, a lysophospholipid, a triacylglyceride, a diglyceride, a glycolipid or a lysoglycolipid and said acyl acceptor is selected from one ore more of the group consisting of a carbohydrate, a protein, a protein subunit, or a hydroxyl acid; and contacting the admixture with a lipid acyltransferase, such that said lipid acyl transferase catalyses one or both of the following reactions: alcoholysis or transesterification.
摘要:
A method for the in situ production of an emulsifier in a foodstuff, wherein a lipid acyltransferase is added to the foodstuff. Preferably the emulsifier is produced without an increase or without a substantial increase in the free fatty acid content of the foodstuff. Preferably, the lipid acyltransferase is one which is capable of transferring an acyl group from a lipid to one or more of the following acyl acceptors: a sterol, a stanol, a carbohydrate, a protein or a sub-unit thereof, glycerol. Preferably, in addition to an emulsifier one or more of a stanol ester or a stanol ester or a protein ester or a carbohydrate ester or a diglyceride or a monoglyceride may be produced. One or more of these may function as an additional emulsifier.