Disclosed is an alcohol dehydrogenase mutant and application thereof in cofactor regeneration, and belongs to the technical fields of enzyme engineering and bioengineering. The alcohol dehydrogenase mutant is obtained by mutating valine at position 84 and/or tyrosine at position 127 in alcohol dehydrogenase having an original amino acid sequence as set forth in SEQ ID No. 1. The alcohol dehydrogenase mutant has high activity for a variety of alcohol co-substrates, and can catalyze these enzyme co-substrates for the regeneration of cofactor NADPH. Compared with the wild-type alcohol dehydrogenase KpADH, the alcohol dehydrogenase mutant has higher activity and catalytic efficiency, and for co-substrate 1,4-butanediol, its k.sub.cat value can be up to 75.9 min.sup.−1, its k.sub.cat/K.sub.m value can be up to 2009 min.sup.−1.Math.M.sup.−1, and its K.sub.m value can be as low as 11.3 mM. Therefore, the alcohol dehydrogenase mutant has a higher value in industrial application.

The disclosure relates, in some aspects, to compositions and methods useful for production of nitrated aromatic molecules. The disclosure is based, in part, on whole cell systems expressing artificial fusion proteins comprising cytochrome P450 enzymes linked to reductase enzymes. In some aspects, the disclosure relates to methods of producing nitrated aromatic molecules in whole cell systems having artificial fusion proteins comprising cytochrome P450 enzymes linked to reductase enzymes.

The present disclosure provides engineered enzymes and methods for preparing tyrosine and tyrosine analogs from a donor amino acid and phenols in a single step. Also disclosed are methods to engineer enzymes having such ‘tyrosine synthase’ activity, starting from the beta-subunit of a tryptophan synthase (TrpB).

The present disclosure provides engineered enzymes and methods for preparing tyrosine and tyrosine analogs from a donor amino acid and phenols in a single step. Also disclosed are methods to engineer enzymes having such ‘tyrosine synthase’ activity, starting from the beta-subunit of a tryptophan synthase (TrpB).

The present invention provides a method for producing an L-amino acid such as a branched-chain L-amino acid by fermentation using a bacterium of the family Enterobacteriaceae, particularly a bacterium belonging to the genus Escherichia, which has been modified to attenuate expression of the gshA gene.

The present invention provides engineered tyrosine ammonia-lyase (TAL) polypeptides and compositions thereof. In some embodiments, the engineered TAL polypeptides have been optimized to provide enhanced catalytic activity and enhanced acid stability, while reducing sensitivity to proteolysis and increasing tolerance to acidic pH levels. The invention also provides methods for utilization of the compositions comprising the engineered TAL polypeptides for therapeutic and industrial purposes.

The present invention provides engineered tyrosine ammonia-lyase (TAL) polypeptides and compositions thereof. In some embodiments, the engineered TAL polypeptides have been optimized to provide enhanced catalytic activity while reducing sensitivity to proteolysis and increasing tolerance to acidic pH levels. The invention also provides methods for utilization of the compositions comprising the engineered TAL polypeptides for therapeutic and industrial purposes.

A polynucleotide, encoding an amino acid sequence, encoding an oxidoreductase, that is ≥50% identical to an amino acid sequence of SEQ ID NO:1 (Geobacillus sp. PA9), SEQ ID NO:3 (Thermus thermophilus), SEQ ID NO:4 (Streptomyces globisporus), SEQ ID NO:5 (Clostridium aminobutyricum), SEQ ID:6 (Burkholderai cepacia), SEQ ID NO:8 (Oscillatoria sp. PCC 6506), or SEQ ID NO:9 (Paraburkholderia phymatum). The polynucleotide has an amino acid exchange in one or more of positions 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214 of SEQ ID NO:1, or at a corresponding position of the amino acid sequence of SEQ ID NO:3, SEQ ID NO:4, SEQ ID:5, SEQ ID NO:6, SEQ ID NO:8, or SEQ ID NO:9.

The present disclosure relates to bacterium engineered to produce aromatic compounds or compounds with aromatic metabolites or intermediates using the CRISPR-CAS transcriptional activation (CRISPRa) and/or transcriptional repression (CRISPRi). Accordingly, in an aspect the present disclosure relates to an engineered bacterium comprising genetic elements supporting programmable transcriptional activation and/or repression. The present disclosure also provides methods and systems for producing aromatic compounds or compounds with aromatic metabolites or intermediates using the engineered bacterium disclosed herein.

The present disclosure relates to a protein variant having a tryptophan-exporting activity, an L-tryptophan-producing microorganism expressing the protein variant, and a method for producing L-tryptophan using the microorganism.