Controlled release antimicrobial sachet prepared from poly(butylene succinate)/geraniol and ethylene vinyl alcohol coated paper for bread shelf-life extension application.

This research aims to develop white bread shelf-life extension sachet with controlled release of antimicrobial agent prepared from multicomponent bio-based materials. The structure of antimicrobial sachet consists of two major parts i.e.

Identification of auxotrophic mutants of the yeast Kluyveromyces marxianus by non-homologous end joining-mediated integrative transformation with genes from Saccharomyces cerevisiae.

The isolation and application of auxotrophic mutants for gene manipulations, such as genetic transformation, mating selection and tetrad analysis, form the basis of yeast genetics. For the development of these genetic methods in the thermotolerant fermentative yeast Kluyveromyces marxianus, we isolated a series of auxotrophic mutants with defects in amino acid or nucleic acid metabolism. To identify the mutated genes, linear DNA fragments of nutrient biosynthetic pathway genes were amplified from Saccharomyces cerevisiae chromosomal DNA and used to directly transform the K.

Designed construction of recombinant DNA at the ura3Δ0 locus in the yeast Saccharomyces cerevisiae.

Recombinant DNAs are traditionally constructed using Escherichia coli plasmids. In the yeast Saccharomyces cerevisiae, chromosomal gene targeting is a common technique, implying that the yeast homologous recombination system could be applied for recombinant DNA construction. In an attempt to use a S.

N-Glycosylation deficiency enhanced heterologous production of a Bacillus licheniformis thermostable α-amylase in Saccharomyces cerevisiae.

Expression of foreign enzymes in yeast is a traditional genetic engineering approach; however, useful secretory enzymes are not produced in every case. The hyperthermostable α-amylase encoded by the AmyL gene of Bacillus licheniformis was expressed in Saccharomyces cerevisiae; however, it was only weakly produced and was degraded by the proteasome. To determine the cause of low α-amylase production, AmyL was expressed in a panel of yeast mutants harboring knockouts in non-essential genes.

Fusion PCR via novel overlap sequences.

Overlap extension or fusion PCR is thought to be a simple and easy method to produce fusion DNA fragments without the need for restriction enzyme digestion and DNA ligation. However, this method has not been used frequently, probably as it is not always reliable. When natural sequences are used for overlap sequences, sometimes either no fusion DNA is produced or only faint DNA bands are detected owing to low annealing between the overlap sequences selected.

Combinatorial gene overexpression and recessive mutant gene introduction in sake yeast.

Industrial yeast strains are generally diploid and are often defective in sporulation. Such strains are hence thought to be less tractable for manipulation by genetic engineering. To facilitate more reliable genetic manipulation of the diploid yeast Japanese sake, we constructed variants of this strain that were homozygous for a URA3 deletion, homozygous for either MATa or MATalpha, and homozygous for either the his3 or the lys4 mutation.

Reliable fusion PCR mediated by GC-rich overlap sequences.

Recombinant DNA technology largely depends upon Escherichia coli plasmid construction via restriction enzyme digestion and DNA ligation. Overlap extension PCR is another simple technique for constructing recombinant DNA but is not commonly used. This is likely due to the inefficiency of fusion after the annealing of overlaps that are generally designed from authentic sequences in the DNA fragments.

High-temperature ethanol fermentation and transformation with linear DNA in the thermotolerant yeast Kluyveromyces marxianus DMKU3-1042.

We demonstrate herein the ability of Kluyveromyces marxianus to be an efficient ethanol producer and host for expressing heterologous proteins as an alternative to Saccharomyces cerevisiae. Growth and ethanol production by strains of K. marxianus and S.