The disclosure relates to a non-natural flavin-dependent oxidase comprising at least one amino acid variation as compared to a wild type flavin-dependent oxidase, wherein the non-natural flavin-dependent oxidase does not comprise a disulfide bond, and wherein the non-natural flavin-dependent oxidase is capable of oxidative cyclization of a prenylated aromatic compound into a cannabinoid. The disclosure also relates to a nucleic acid, an expression construct, and an engineered cell for making the non-natural flavin-dependent oxidase. Also provided are compositions comprising the non-natural flavin-dependent oxidase; isolated non-natural flavin-dependent oxidase and methods of making the same; cell extracts comprising the non-natural flavin-dependent oxidase; and methods of making cannabinoids.

The present invention relates to a recombinant strain for producing shikimic acid, in which a target gene that regulates the asymmetric cell division and target genes that regulate the shikimic acid production are expressed The target gene that regulates the asymmetric cell division includes cytoskeletal protein PopZ coding gene popZ, and the target genes that regulate the shikimic acid production include DAHP synthase coding gene aroG, 3-dehydroquinate synthase coding gene aroB, and transketolase coding gene tktA. The recombinant strain of the present invention realizes the de novo synthesis of shikimic acid using glucose as a substrate, with a low cost. After fermentation with the strain in a 7.5 L fermentor, the highest production of shikimic acid is 88.1 g/L, the yield is 0.33 g/g, and the production intensity of shikimic acid is 1.1 g/L/h.

Provided is a nucleic acid comprising a sequence encoding a prenyltransferase (PT) gene or its complement, codon optimized for production in a microorganism or a plant. Also provided is a yeast expression cassette comprising the above nucleic acid. Additionally provided is a non-naturally occurring prenyltransferase (PT) comprising an amino acid sequence having at least 90% amino acid sequence identity or conservative amino acid substitutions to the amino acid sequences encoded by the above nucleic acid. Further provided is a recombinant microorganism or plant expressing a PT encoded by the above nucleic acid. Additionally provided is a method of catalyzing the condensation of a polyprenol diphosphate and an alkylresorcinol or alkylresorcyclic acid to yield a cannabinoid.

The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of metabolically engineered cells and use of the cells in a cultivation or fermentation. The disclosure describes a metabolically engineered cell and a method by cultivation or fermentation with the cell for production of a sialylated di- and/or oligosaccharide. The metabolically engineered cell comprises a pathway for production of the sialylated di- and/or oligosaccharide and is modified for expression and/or overexpression of multiple coding DNA sequences encoding one or more isoproteins that catalyze the same chemical reaction. Furthermore, the disclosure provides for purification of the sialylated di- and/or oligosaccharide from the cultivation.

The present invention relates to a process for synthesizing functionalized mercaptans essentially in the absence of oxygen, and also to a composition making it possible in particular to implement this process. Said functionalized mercaptans are of the following formula (I):

##STR00001##

in which, R.sub.1 and R.sub.7, which are identical or different, are a hydrogen atom or an aromatic or nonaromatic, linear, branched or cyclic, saturated or unsaturated, hydrocarbon chain of 1 to 20 carbon atoms which may comprise one or more heteroatoms; X is chosen from -C(=O)-, -CH.sub.2- or -CN; R.sub.2 is: (i) either absent when X represents -CN, (ii) or a hydrogen atom, (iii) or -OR.sub.3, R.sub.3 being a hydrogen atom or an aromatic or nonaromatic, linear, branched or cyclic, saturated or unsaturated, hydrocarbon chain of 1 to 20 carbon atoms which may comprise one or more heteroatoms, (iv) or -NR.sub.4R.sub.5, R.sub.4 and R.sub.5, which are identical or different, being a hydrogen atom or an aromatic or nonaromatic, linear, branched or cyclic, saturated or unsaturated, hydrocarbon chain of 1 to 20 carbon atoms which may comprise one or more heteroatoms; n is equal to 1 or 2; and * represents an asymmetric carbon.

The present invention describes methods to produce a synthetic or semi-synthetic cysteine synthase/PLP-dependent decarboxylase (sCs/PLP-DC). More particularly, the invention relates to genetic modification of organisms including eukaryotes and prokaryotes to express a functional sCs/PLP-DC. The invention includes methods to produce taurine in organisms that contain native or heterologous (transgenic) taurine biosynthetic pathways or cells that have taurine by enrichment. The invention also relates to methods to increase taurine levels in the cells and to use the said cells or extracts or purifications from the cells that contain the invention to produce plant growth enhancers, food, animal feed, aquafeed, food or drink supplements, animal-feed supplements, dietary supplements, health supplements or taurine.

Provided herein are programmable condensate protein systems and nucleic acid constructs encoding the same. The protein system enables modular targeting of proteins of interest. Protein-peptide interaction domains (PPIDs) are incorporated to functionalize engineered condensates with the attributes of the recruited protein, resulting in a modular system that allows for diverse facile and reprogrammable applications, including in enzyme clustering of metabolic pathways. Colocalizing specific metabolic enzymes in these condensates results in functionalized organelles with which can be used to manipulate the output of engineered metabolic pathways for the production of a pharmaceutical precursor.

Methods for recombinant production of steviol glycoside and compositions containing steviol glycosides are provided by this invention.

The invention relates generally to materials and methods for biosynthesising quillaic acid in a host by expressing heterologous nucleotide sequences in the host each of which encodes a polypeptide which in combination have said QA biosynthesis activity. Example polypeptides include (i) a Beta-amyrin synthase; (ii) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-28 position to a carboxylic acid; (iii) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-16a position to an alcohol; and (iv) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-23 position to an aldehyde. Preferred nucleotide sequences are obtained from, or derived from, Q. saponaria.

The present invention relates to a biocatalytical method for manufacturing of homoeriodictyol dihydrochalcone and/or hesperetin dihydrochalcone by providing at least one first biocatalyst system for the hydroxylation of phloretin and/or its glycosides as well as at least one second biocatalyst for the methylation of 3-hydroxyphloretin. Further disclosed are microorganisms capable of producing such biocatalysts as well as sequences encoding the biocatalysts. Furthermore, the present invention relates to the use of a mixture obtained by a method as disclosed in the present invention and to specific compositions suitable as sweetness enhancers and/or flavouring agents.