The present invention relates to isolated polypeptides having cellobiohydrolase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Provided herein is a genetically engineered microorganism comprising knock-in of DNA at an acetolactate decarboxylase gene locus. Replacement of the acetolactate decarboxylase gene with DNA encoding one or more native or nonnative enzymes confers certain advantages, including fermentation stability and increased production of native and nonnative products from gaseous substrates.

Provided herein is a genetically engineered microorganism comprising knock-in of DNA at an acetolactate decarboxylase gene locus. Replacement of the acetolactate decarboxylase gene with DNA encoding one or more native or nonnative enzymes confers certain advantages, including fermentation stability and increased production of native and nonnative products from gaseous substrates.

This disclosure generally relates to the use of enzyme combinations or recombinant microbes comprising same to make isoprenoid precursors, isoprenoids and derivatives thereof including prenylated aromatic compounds. Novel metabolic pathways exploiting Claisen, aldol, and acyloin condensations are used instead of the natural mevalonate (MVA) pathway or 1-deoxy-d-xylulose 5-phosphate (DXP) pathways for generating isoprenoid precursors such as isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), and geranyl pyrophosphate (GPP). These pathways have the potential for better carbon and or energy efficiency than native pathways. Both decarboxylative and non-carboxylative condensations are utilized, enabling product synthesis from a number of different starting compounds. These condensation reactions serve as a platform for the synthesis of isoprenoid precursors when utilized in combination with a variety of metabolic pathways and enzymes for carbon rearrangement and the addition/removal of functional groups. Isoprenoid alcohols are key intermediary products for the production of isoprenoid precursors in these novel synthetic metabolic pathways. These precursors can be modified to various isoprenoid products through prenyl transferase, terpene synthase, or terpene cyclases. The production of prenylated aromatic compounds is achieved through prenyl transfer of the hydrocarbon units of isoprenoid precursors to polyketides.

The present invention relates to a biocatalytic process for producing an aroma substance or a mixture of aroma substances, comprising the steps of: providing a conversion medium comprising a plant component of the gooseber-ry family, the rose family and/or the grapevine family; contacting the conversion medium with at least one fungus from the division of stander fungi capable of forming an aroma substance or a mixture of aroma sub-stances on the conversion medium; converting the plant component to the aroma substance or mixture of aroma substances with the aid of the fungus; and optionally recovering the aroma substance or mixture of aroma substances, wherein the aroma substance or mixture of aroma substances preferably com-prises at least one compound selected from the group consisting of 2-octanone, 2-nonanone, 2-undecanone, linalool oxides, benzaldehyde, geraniol, 2-octanol, methylanthranilate, 2-aminobenzaldehyde and linalool.

The present invention relates to a biocatalytic process for producing an aroma substance or a mixture of aroma substances, comprising the steps of: providing a conversion medium comprising a plant component of the gooseber-ry family, the rose family and/or the grapevine family; contacting the conversion medium with at least one fungus from the division of stander fungi capable of forming an aroma substance or a mixture of aroma sub-stances on the conversion medium; converting the plant component to the aroma substance or mixture of aroma substances with the aid of the fungus; and optionally recovering the aroma substance or mixture of aroma substances, wherein the aroma substance or mixture of aroma substances preferably com-prises at least one compound selected from the group consisting of 2-octanone, 2-nonanone, 2-undecanone, linalool oxides, benzaldehyde, geraniol, 2-octanol, methylanthranilate, 2-aminobenzaldehyde and linalool.

The present invention describes an oligomer for use as a UV stabiliser. In particular, the oligomer is suitable for use as a UV stabiliser in a polymer matrix. The present invention also describes a method of forming said oligomer. The method of forming said oligomer comprises a polymerising step, wherein the polymerising step comprises forming a C—C bond on the hydroxyphenyl ring of a monomer. In preferred embodiments, the oligomer is formed from polymerizing bio-derived monomer such as curcumin, its hydrogenated analogue, and an aldol condensation product of cyclic ketone and vanillin.

The present invention describes an oligomer for use as a UV stabiliser. In particular, the oligomer is suitable for use as a UV stabiliser in a polymer matrix. The present invention also describes a method of forming said oligomer. The method of forming said oligomer comprises a polymerising step, wherein the polymerising step comprises forming a C—C bond on the hydroxyphenyl ring of a monomer. In preferred embodiments, the oligomer is formed from polymerizing bio-derived monomer such as curcumin, its hydrogenated analogue, and an aldol condensation product of cyclic ketone and vanillin.

The present invention discloses a ω-transaminase mutant obtained through DNA synthetic shuffling combined mutation. The ω-transaminase mutant is obtained through point mutation of a wild type ω-transaminase from Aspergillus terrus. The amino acid sequence of the wild type ω-transaminase is shown in SEQ ID NO: 1. The mutation site of the ω-transaminase mutant is any one of: (1) F115L-H210N-M150C-M280C; (2) F115L-H210N; (3) F115L-H210N-E253A-I295V; (4) I77L-F115L-E133A-H210N-N245D; (5) I77L-Q97E-F115L-L118T-E253A-G292D; (6) I77L-E133A-N245D-G292D; and (7) H210N-N245D-E253A-G292D. According to the present invention, forward mutations obtained in the previous stage are randomly combined through a DNA synthetic shuffling combined mutation method. It is verified through experiments that this method can effectively improve the probability of forward mutation and increase experimental efficiency and feasibility, and is capable of obtaining mutant enzymes with thermodynamic stability remarkably superior to that of wild enzymes via screening.

The present invention discloses a ω-transaminase mutant obtained through DNA synthetic shuffling combined mutation. The ω-transaminase mutant is obtained through point mutation of a wild type ω-transaminase from Aspergillus terrus. The amino acid sequence of the wild type ω-transaminase is shown in SEQ ID NO: 1. The mutation site of the ω-transaminase mutant is any one of: (1) F115L-H210N-M150C-M280C; (2) F115L-H210N; (3) F115L-H210N-E253A-I295V; (4) I77L-F115L-E133A-H210N-N245D; (5) I77L-Q97E-F115L-L118T-E253A-G292D; (6) I77L-E133A-N245D-G292D; and (7) H210N-N245D-E253A-G292D. According to the present invention, forward mutations obtained in the previous stage are randomly combined through a DNA synthetic shuffling combined mutation method. It is verified through experiments that this method can effectively improve the probability of forward mutation and increase experimental efficiency and feasibility, and is capable of obtaining mutant enzymes with thermodynamic stability remarkably superior to that of wild enzymes via screening.