This invention is an improved method of robust and scalable production of precursors of active cannabinoids, including geranyl pyrophosphate (GPP) and/or cannabigerolic acid (CBGA), in a methylotrophic yeast host cell. The improved methods incorporate a polypeptide encoding an Erg20 variant (F98W/N128W) into a methylotrophic yeast host cell, for example Pichia pastoris (Komagataella phaffii), that biases the natural production of FPP and GPP towards GPP, a precursor to the intermediate CBGA, crucial to the synthesis of active cannabinoids.

The present disclosure relates to automated multi-module instruments, compositions and methods for performing nucleic acid-guided nuclease editing; specifically, methods, instruments, systems, and nucleic acids synthetic cassettes that improve the efficiency of gene editing using CRISPR enzymes in diploid cells—either on both chromosomes or selectively in one chromosome—without incurring loss of heterozygosity (LOH) and its often deleterious effects.

The present invention is directed to compositions comprising hydrolase-secreting bacteria and fermenting microorganisms and use thereof, such as for fermentative production of ethanol.

The present disclosure provides methods for producing consumable recombinant proteins that are substantially free from herein-disclosed undesired byproducts.

A recombinant human type XVII collagen consists of an amino acid sequence shown in (A).sub.n or includes the amino acid sequence shown in (A).sub.n, where A is a sequence set forth in SEQ ID NO: 2, a sequence undergoing an amino acid modification to a predetermined extent based on SEQ ID NO: 2, or a sequence that has more than 80% homology with SEQ ID NO: 2; n is an integer greater than or equal to 1; and A represents a basic unit, and when there are two or more basic units, the two or more basic units are identical or different and are directly connected in tandem through a peptide bond. In the present disclosure, it is confirmed that the recombinant human type XVII collagen can undergo efficient secretory and soluble expression in eukaryotic host cells such as Pichia pastoris (P. pastoris).

The present application describes improved yeast culture media that are particularly suitable for cultivation of Pichia yeast, in particular P. kluyveri. Also described herein are methods of cultivating yeast using this improved culture medium. Further described herein are improved methods of producing a fermented juice beverage.

A novel gene targeting method and a nucleotide construct for the method. The method integrates a nucleotide construct containing an interference gene in an effective gene targeting region independent of the gene by homologous recombination, thereby improving the targeting efficiency of the gene. The present invention also provides a gene targeting system for gene expression regulation and gene disruption.

The present invention relates to a method for producing a recombinant protein in yeast cells, wherein the cells are subjected to a temperature shift at a specific timepoint of the cell culture. It also relates to a method for producing a recombinant protein in yeast cells by culturing said yeast cells in a medium having a high concentration of potassium ions compared to the concentration of sulfate and/or phosphate ions.

The biological production of beta-hydroxyisovalerate (βHIV) using a non-natural microorganism. The non-natural microorganism for the biologically-derived βHIV provides more beta-hydroxyisovalerate synthase activity than the wild-type parent. The non-natural microorganism can host a non-natural enzyme, such as the non-natural enzyme expressed in a yeast or bacteria, wherein the non-natural microorganism comprises an active βHIV metabolic pathway for the production of βHIV. The biological derivation of βHIV eliminates toxic by-products and impurities that result from the chemical production of βHIV, such that βHIV produced by a non-natural microorganism prior to any isolation or purification process has not been in substantial contact with any halogen-containing component.

The disclosure discloses a genetically engineered strain for producing porcine myoglobin and fermentation and purification thereof, and belongs to the technical field of genetic engineering. The disclosure realizes efficient secretion and expression of porcine myoglobin by integrating the gene of porcine myoglobin in P. pastoris. On this basis, optimization of the medium and culture conditions of recombinant P. pastoris can significantly increase the titer of porcine myoglobin, so that the titer can reach 285.42 mg/L under fermenter conditions. In addition, by creatively adding different concentrations of ammonium sulfate to fermentation broth step by step, the purity of myoglobin obtained by final concentration is up to 88.0%, and the purification rate is up to 66.1%. The disclosure realizes efficient expression and high purification of porcine myoglobin from various steps such as synthesis, fermentation and purification of porcine myoglobin, and provides broad prospects for industrial production of porcine myoglobin.