The present disclosure application relates to a construction method of a yeast display library (YSD), specifically to a construct for a yeast display library, an expression vector, a host cell and a construction method and use thereof. The yeast display library provided by the present application has high transformation efficiency and rich diversity.

Characterization of the binding dynamics at the interface between any two proteins that specifically interact plays a role in myriad biomedical applications. The methods disclosed herein provide for the high-throughput characterization of the specific interaction at the interface between two protein binding partners and the identification of functionally significant mutations of one or both protein binding partners. For example, the methods disclosed herein may be useful for epitope and paratope mapping of an antibody-antigen pair, which is useful for the discovery and development of novel therapies, vaccines, diagnostics, among other biomedical applications.

The present disclosure provides a method for genetically engineering Yarrowia lipolytica host cell for producing glycolic acid from organic wastes. A subject genetically engineered Y. lipolytica cell comprises the disrupted native genes encoding malate synthase, heterologous enzyme of glyoxylate reductase targeted in the different cellular compartments including mitochondria, peroxisome and cytosol, and a mutant NADP.sup.+-dependent malate dehydrogenase. The pathway with a theoretical yield as high as that 1 g of acetic acid can be converted to 1.27 g of glycolic acid without carbon loss was engineered for glycolic acid production. The methods particularly include process for production of volatile fatty acids (VFAs) mainly comprised of acetic acid from organic waste, and then use of resultant VFAs for biosynthesis of glycolic acid by recombinant Y. lipolytica.

Compositions, devices, and systems for use in a high-throughput screening platform for identifying anti-aging compounds and/or mutations that extend replicative life span (RLS). Specifically, herein disclosed is a yeast cell daughter-arresting-program (DAP), as well as compositions used in devices and systems that allow measurement of replicative lifespan and identification of agents or mutations that modulate the lifespan.

Provided herein are genetically modified Issatchenkia yeast and fermentation methods for producing succinic acid.

A method for producing an objective substance such as phytosphingosine (PHS) and phytoceramide (PHC) using yeast is provided. The objective substance is produced by cultivating yeast having an ability to produce the objective substance and modified so that the expression and/or activity of a protein(s) encoded by NEM1 and/or SPO7 gene(s) is reduced in a culture medium.

Characterization of the binding dynamics at the interface between any two proteins that specifically interact plays a role in myriad biomedical applications. The methods disclosed herein provide for the high-throughput characterization of the specific interaction at the interface between two protein binding partners and the identification of functionally significant mutations of one or both protein binding partners. For example, the methods disclosed herein may be useful for epitope and paratope mapping of an antibody-antigen pair, which is useful for the discovery and development of novel therapies, vaccines, diagnostics, among other biomedical applications.

Disclosed is a method for manufacturing a monoclonal antibody without using animal individuals. This method includes a step of introducing a DNA fragment comprising a gene that encodes a secretory signal, a gene that encodes a nanobody, and a gene that encodes a peptide barcode, or a vector containing the DNA fragment, into a yeast cell; and a step of collecting a polypeptide comprising the nanobody and the peptide barcode that has been expressed in the cell and secreted to the outside of the cell. According to the method, it is possible to manufacture a monoclonal nanobody more efficiently in a shorter period of time without using animal individuals.

The present invention discloses a construction method and a recombinant yeast stain Yarrowia lipolytica for xylitol synthesis; Adopting Yarrowia lipolytica as the host, introducing genes into the host through metabolic engineering to enable the recombinant yeast to synthesize xylitol from glucose, fructose, glycerol and starch as carbon sources, block the synthesis pathway of by-products, so that it can synthesize xylitol from the aforesaid carbon sources by fermentation, thus obtain the engineered Yarrowia lipolytica strain to synthesize xylitol from glucose and other carbon sources. After fermentation, xylitol crystal is obtained by ion exchange, decolorization, concentration and crystallization of the clear and transparent fermentation liquor after isolation of the strains from the fermentation. This construction method of engineered Yarrowia lipolytica described in the invention, and the Yarrowia lipolytica strain obtained by this method can simplify the existing method for chemical synthesis of xylitol and have good application.

A recombinant yeast is constructed by introducing an expressed gene of exogenous Fructose-6-phosphate phosphoketolase into a modified yeast cell, and the modified yeast cell is a yeast cell with a metabolic pathway for synthesizing tyrosol via Erythrose-4-phosphate and phosphoenolpyruvate. The present invention discloses for the first time that in the process of expressing Fructose-6-phosphate phosphoketolase in a yeast, Fructose-6-phosphate is synthesized into beta-D-Fructose 1,6-bisphosphate and also catalyzed into Erythrose-4-phosphate and Acetyl-phosphate, and Xylulose-5-phosphate is catalyzed into Glyceraldehydes-3-phosphate and Acetyl-phosphate, which change the metabolic flux distribution of carbon in the yeast, enhance the synthesis of Erythrose-4-phosphate as an important intermediate for the biosynthesis of tyrosol, optimize the metabolic pathway for synthesizing tyrosol, and increase the yields of tyrosol and its derivatives such as hydroxytyrosol.