Cibt America Inc.

Cathay Biotech Inc.

Cathay R&d Center Co., Ltd

Cathay Biotec Inc.

Howard Chou

Xiucai Liu

Min Xu

Ling Chen

Wenbo Liu

Chen Yang

Wenqiang Lu

Naiqiang Li

Lijun Yu

Yunfeng Lei

This invention provides acid decarboxylase-prion subunit fusion polypeptides, nucleic acid sequences, expression vectors, and host cells expression such fusion polypeptides to produce various amino acids and derivatives of the amino acids such as polyamines.

Method of improving acid decarboxylase activity in vitro under alkaline pH

The present invention relates to a long-chain dibasic acid with low content of long-chain dibasic acid impurity of shorter carbon chain, to the preparation of a long-chain dibasic acid producing strain by directed evolution of POX gene and homologous recombination, and to the production of a long-chain dibasic acid with low content of long-chain dibasic acid impurity of shorter carbon chain by using the strain. The present invention also relates to a strain containing a mutated promoter, wherein, when a long-chain dibasic acid is produced by fermentation of this strain, the content of the acid impurity of shorter carbon chain in the fermentation product is significantly reduced.

Long chain dibasic acid with low content of long chain dibasic acid impurity of shorter carbon-chain and preparation method thereof

Provided microorganisms genetically modified to overexpress an imine/enamine deaminase to enhance the production of lysine and lysine derivatives by the microorganism. Also provided a method of generating such microorganism, and methods of producing lysine and lysine derivatives using the genetically modified microorganisms.

Reducing the accumulation of imines/enamines for the production of amino acids or amino acid-derived products

The invention is directed to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution and homologous recombination, a strain obtained by this method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use of the strain. In particular, the invention is directed to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution of CYP52A12 gene and homologous recombination, a strain obtained by this method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use of the strain.

Directed evolution of CYP52A12 gene and its use in dicarboxylic acid production

The invention provides CadA polypeptides with mutations that increase activity in alkaline pH compared to the wild-type lysine decarboxylase. The invention also provides methods of generating such mutant polypeptides, microorganisms genetically modified to overexpress the mutant polypeptides, and methods of generating such microorganism.

Modifications to lysine decarboxylase enzymes

The invention provides microorganisms genetically modified to overexpress biofilm dispersal related polypeptides to enhance the production of lysine and lysine derivatives by the microorganism, method of generating such microorganism, and methods of producing lysine and lysine derivatives using the genetically modified microorganisms.

Cells and methods for production of lysine and lysine-derived products

One aspect provided herein relates to lysine decarboxylase polypeptides comprising mutants of SEQ ID NO: 2 (i.e., mutants of() Ldc) and/or fragments thereof. In certain embodiments, the mutants or fragments thereof may have at least about 95% sequence identity with SEQ ID NO: 2. Another aspect provided herein relates to a DNA polynucleotide comprising one or more lysine decarboxylase nucleotide sequences of mutants of SEQ ID NO: 1 (i.e., mutants ofldc), fragments thereof, or fragments of SEQ ID NO: 1 (i.e., fragments ofldc). In certain embodiments, the DNA polynucleotide as disclosed herein may encode one or more lysine decarboxylase polypeptides provided herein. In certain embodiments, the DNA polynucleotide as disclosed herein may encode SEQ ID NO: 2, mutants, and/or fragments thereof. In certain embodiments, the lysine decarboxylase nucleotide sequences provided herein may have at least 95% sequence identity with SEQ ID NO: 1 or SEQ ID NO: 3. Another aspect provided herein relates to expression vectors comprising the DNA polynucleotides described herein used for production of a lysine-derived product. Other aspects provided herein include transformants, mutant host cells, methods for the production of lysine decarboxylases, and methods for the production of a lysine-derived product.

Expression ofpolypeptides involved in lysine decarboxylation, and methods and applications thereof

The invention relates to a long-chain dibasic acid with low content of monobasic acid impurity and a production method thereof, in particular to the preparation of a long-chain dibasic acid producing strain by means of directed evolution and homologous recombination, and to the production of a long-chain dibasic acid with low content of monobasic acid impurity by fermentation of said strain. The invention relates to a mutated CYP52A12 gene, homologous gene or variant thereof, which, relative to GenBank Accession Number AY230498 and taking the first base upstream of the start codon ATG as &#x2212;1, comprises a mutation. The invention relates to a strain comprising said mutated CYP52A12 gene, homologous gene or variant thereof wherein when the strain is fermented to produce a long-chain dibasic acid, the content of monobasic acid impurity in the fermentation product is significantly reduced.

Long-chain dibasic acid with low content of monobasic acid impurity and the production method thereof

The invention provides microorganisms genetically modified to overexpress thermophilic lysine decarboxylase polypeptides in a mesophilic host to enhance the production of lysine and lysine derivatives by the microorganism, method of generating such microorganism, and methods of producing lysine and lysine derivatives using the genetically modified microorganisms.

Heterologous expression of thermophilic lysine decarboxylase and uses thereof

The present invention relates to a long-chain dibasic acid with low content of hydroxyl acid impurity and a production method thereof, in particular to a method for producing a long-chain dibasic acid with low content of hydroxyl acid impurity by fermenting a long-chain dibasic acid producing strain prepared by homologous recombination method. The present invention relates to a recombinant long-chain dibasic acid producing microorganism, having increased alcohol dehydrogenase activity and optionally decreased acyl-CoA oxidase activity. The present invention also relates to a method of producing a long-chain dibasic acid by the recombinant long-chain dibasic acid producing microorganism and use thereof.

Long-chain dibasic acid with low content of hydroxyl acid impurity and production method thereof

The invention provides microorganisms genetically modified to co-overexpress a carbohydrate binding module and lysine decarboxylase polypeptides in a mesophilic host to enhance the production of lysine derivatives by the microorganism, method of generating such microorganism, and methods of producing lysine derivatives using the genetically modified microorganisms.

Heterologous expression of carbohydrate binding modules and uses thereof for cadaverine production

One aspect of the invention relates to a mutant polypeptide comprising the amino acid sequence oftetracycline efflux pump A (TetA). Another aspect of the invention relates to a polynucleotide encoding a polypeptide of TetA, a fragment thereof, or a mutant thereof. Another aspect of the invention relates to a first expression plasmid vector comprising one or more first polynucleotides encoding a first polypeptide comprising a tetracycline efflux pump polypeptide, a fragment thereof or a mutant thereof, one or more second polynucleotides independently selected from the group consisting of a third polynucleotide encoding a third polypeptide comprising a lysine decarboxylase polypeptide, a fragment thereof or a mutant thereof, and a fourth polynucleotide encoding a fourth polypeptide comprising a lysine biosynthesis polypeptide, a fragment thereof or a mutant thereof. Another aspect of the invention relates to a transformant comprising one or more expression plasmid vectors as described herein in a host cell. Another aspect of the invention relates to a mutant host cell comprising one or more first, third or fourth polynucleotides as described herein integrated into a chromosome of a host cell. Another aspect of the invention relates to a method for producing a lysine comprising obtaining a transformant and/or mutant host cell as disclosed herein, culturing the transformant and/or mutant host cell under conditions effective for the expression of the lysine; and harvesting the lysine. Other aspects of the invention relate to methods for producing biobased cadaverine using the transformants disclosed herein, and biobased cadaverine prepared by the method disclosed herein, and polyamides formed using the biobased cadaverine as disclosed herein and compositions thereof.

Expression of recombinant tetracycline efflux pumps for the production of lysine or lysine-derived products, and methods and applications thereof

The invention relates to the directed evolution of CYP52A12 gene and the use thereof for the production of a dicarboxylic acid. In particular, it relates to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution and homologous recombination, a strain obtained by the method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use thereof. In particular, the invention relates to a method of preparing a long chain dicarboxylic acid producing strain by directed evolution of CYP52A12 gene and homologous recombination, a strain obtained by the method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use thereof. By directed evolution of CYP52A12 gene, one strain which has a base mutation at the promoter region of said gene and is capable of producing a long chain dicarboxylic acid under an acidic condition in a shortened fermentation time is screened out in the invention.

The invention provides CadA polypeptides with mutations that increase activity in alkaline pH compared to the wild-type lysine decarboxylase. The invention also provides methods of generating such mutant polypeptides, microorganisms genetically modified to overexpress the mutant polypeptides, and methods of generating such microorganisms.

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Shanghai, China

Howard Chou