A process of recovering oil, comprising (a) converting a starch-containing material into dextrins with an alpha-amylase; (b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar; (c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism; (d) recovering the fermentation product to form a whole stillage; (e) separating the whole stillage into thin stillage and wet cake; (e) optionally concentrating the thin stillage into syrup; (f) recovering oil from the thin stillage and/or optionally the syrup, wherein a protease and a phospholipase are present and/or added during steps (a) to (c). Use of a protease and a phospholipase for increasing oil recovery yields from thin stillage and/or syrup in a fermentation product production process.

The present application relates to the field of enzyme engineering, especially relates to a pullulanase as well as preparation and use thereof. The pullulanase and coding gene thereof were obtained by random mutation by using the Error-prone PCR technique on the gene of wild-type pullulanase to obtain a mutant PLUM. The enzyme activity of the mutant PLUM was improved by 57.03% compared with the wild-type pullulanase PLUM.

A method of preparing a wort with an increased level of free amino nitrogen (FAN) comprising: a) preparing a mash from a grist comprising malt and/or adjunct; and b) adding a protease having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1.

The present invention relates to the field of genetic engineering, particularly to a method for efficiently expressing pullulanase in Bacillus subtilis and recombinant Bacillus subtilis. said method includes steps of constructing modified Bacillus subtilis strain with deletion of alkaline protease gene and neutral protease gene, constructing expression vector including an optimized combination of promoter and signal peptide and pullulanase gene, and transforming said modified Bacillus subtilis strain with by said expression vector. A series of combinations of promoter and signal peptide are optimized to obtain the combination for efficiently expressing pululanase, provide an industrial application basis.

A process of recovering oil, comprising (a) converting a starch-containing material into dextrins with an alpha-amylase; (b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar; (c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism; (d) recovering the fermentation product to form a whole stillage; (e) separating the whole stillage into thin stillage and wet cake; (e) optionally concentrating the thin stillage into syrup; (f) recovering oil from the thin stillage and/or optionally the syrup, wherein a protease and a phospholipase are present and/or added during steps (a) to (c). Use of a protease and a phospholipase for increasing oil recovery yields from thin stillage and/or syrup in a fermentation product production process.

The present disclosure discloses a maltooligosyl trehalose synthase mutant with improved thermal stability, and belongs to the technical fields of enzyme engineering and protein engineering. The residual enzyme activities of the MTSase mutants S361R, S444E, S361R/S444E, S361K/S444E, G415P/S361R/S444E and G415P consistent with the present disclosure after treatment at 60 C. for 10 min are respectively 70.3%, 50.1%, 83.5%, 65.9%, 100% and 80.7%, which are respectively 1.6, 1.1, 1.9, 1.5, 2.3 and 1.9 times of that of the wild type. The half-lives of the S361R/S444E and G415P/S361R/S444E at 60 C. are respectively 14.9 min and 90.8 min which are respectively 3.2 and 19.7 times of that of the wild type, indicating that the thermal stability of the MTSase mutant consistent with the present disclosure is significantly improved than that of the wild type.

Methods of using thermostable serine proteases are described herein.

The present disclosure relates to a method for preparing myo-inositol using myo-inositol monophosphate synthase consisting of an amino acid sequence of SEQ ID NO: 1 and/or myo-inositol monophosphate phosphatase consisting of an amino acid sequence of SEQ ID NO: 3.

A method of preparing a wort with an increased level of free amino nitrogen (FAN) comprising: a) preparing a mash from a grist comprising malt and/or adjunct; and b) adding a M4 mettalloprotease obtainable from Actinobacteria; and wherein the amount of free amino nitrogen (FAN) in the wort is increased as compared to a wort produced in the absence of the M4 metalloprotease.

Starch spherulites are produced by debranching of amylopectin-containing starch into short linear -1,4-linked glucans (e.g., short-chain amylose, SCA). The debranched linear glucans are directly converted into spherulites by heating the debranched starch mixture followed by cooling and crystallization to form well-developed spherulites. The spherulites exhibit controlled enzyme digestibility.