The disclosure provides biosynthetic platforms that generate olivetolic acid and its analogues at high titers from microbes, and in cell free systems.

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 methods for identifying binding partners using cell surface display libraries, where the cells of the library display engineered peptides on their cell surfaces for identification of peptides that bind to targets of interest. The engineered peptides are preferably expressed in the cells under conditions that provide both secretion and display of the engineered peptides on the cell surfaces, thus providing access of the engineered peptides to identify potential binding pairs. The cell libraries cab be engineered using an automated editing system that provides for one or more targeted edits per cell.

Isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are disclosed. More specifically, nucleic acids isolated from Pichia pastons having promoter activity and expression methods, host cells, expression vectors, and DNA constructs of using the Pichia pastons promoters to produce different proteins and polypeptides are disclosed.

The invention discloses the application of a β-N-acetylhexosaminidase (HaHex74) from Haloferula sp. in the synthesis of human milk oligosaccharides. The invention provides the use of HaHex74 protein or related biological materials thereof in any one of the following: synthesizing human milk oligosaccharides; synthesizing Lacto-N-triose II and/or Lacto-N-neotetraose; the HaHex74 protein having the amino acid sequence shown in SEQ ID No. 2 is derived from Haloferula sp. The β-N-acetylhexosaminidase HaHex74 disclosed by the invention possesses high-level expression, excellent hydrolysis properties and transglycosylation activity, which may make it potentially useful in the production of human milk oligosaccharides.

Provided herein are compositions, proteins, polynucleotides, expression vectors, host cells, kits, and systems for producing egg white proteins, as well as methods of using the same.

According to the present disclosure, on the basis of all existing mutations, the tenth glycine of a BC-PC-PLC is mutated into aspartic acid, a specific enzyme activity thereof is 83% higher than that of a sequence before the mutation, and protein expression and degumming activity of unit enzyme activity do not change, so as to further reduce manufacturing costs.

The present invention refers to the heterologous production in microorganisms of modified versions of a predicted isoform of the surfactant protein Lv-ranaspumin-1 (Lv-Rsn-1), whose sequence was inferred from analyzes of the protein extract of the nest foam from the Northeastern Pepper Frog (Leptodactylus vastus). More specifically, it refers to two surfactant proteins that consist of modified versions of the predicted isoform of Lv-Rsn-1; to two synthetic genes each encoding one of these modified versions of the predicted isoform of Lv-Rsn-1; to two expression cassettes each containing one of the synthetic genes encoding one of the modified versions of the predicted isoform of Lv-Rsn-1; to two expression vectors each containing one of the synthetic genes encoding modified versions of the predicted isoform of Lv-Rsn-1; and to two transgenic microorganisms, a bacterium and a yeast, each transformed with one of these synthetic genes and heterologously producing one of the modified versions of the predicted isoform of Lv-Rsn-1. Lv-Rsn-1 has surfactancy, emulsification and dispersancy properties, among others, and its heterologous production allows it to be used in various applications and industrial products, without the need to extract it from the frog nest foam.

Provided herein are compositions with enzymatically active enzymes produced recombinantly, enhanced protein content and methods for the preparation thereof.

The present invention provides nucleic acids, vectors, host cells and methods for production of beta-fructofuranosidase from Aspergillus niger. The invention represents an advancement in the field of genetic engineering and provides methods for obtaining high yield of a novel recombinant β-fructofuranosidase encoded by fopA gene of Aspergillus niger as a secreted protein.