Two distinct hemolysins were demonstrated in Vibrio parahaemolyticus. A thermostable direct hemolysin purified from V. parahemolyticus WP-1, a Kanagawa phenomenon (KP)-positive strain, is antigenically different from a thermolabile hemolysin produced by V. parahaemolyticus T-3454, a KP-negative strain. The thermostable direct hemolysin was found in KP-positive strains but not in KP-negative str...

A hyperthermophilic archaeon Pyrococcus horikoshii was isolated from a hydrothermal vent in the Okinawa trough in the Pacific Ocean. It optimally grows at around 100oC. The hyperthermophiles were expected to be good producers of thermostable enzymes active at around 100 oC. The thermostable enzymes have considIkuo MATSUI Biological Information Research Center e-mail: [email protected] AIST T...

We describe a method to genetically manipulate Chaetomium thermophilum, a eukaryotic thermophile, along with various biochemical applications. The transformation method depends on a thermostable endogenous selection marker operating at high temperatures combined with chromosomal integration of target genes. Our technique allows exploiting eukaryotic thermophiles as source for purifying thermost...

Changes in the soluble protein content, electrophoretic profiles of total and thermostable proteins and polypeptides in endosperms and roots of wheat seedlings germinated under optimal and stress conditions were investigated. Low and high temperature, 0.2 M NaCl, 0.5 M sucrose, 30 μM ABA and 10 mM H2O2 were applied as stress factors and seeds germinated for 72 hours. The results obtained show t...

The ability to design thermostable proteins offers enormous potential for the development of novel protein bioreagents. In this work, a combined computational and experimental method was developed to increase the T m of the flavin mononucleotide based fluorescent protein Bacillus Subtilis YtvA LOV domain by 31 Celsius, thus extending its applicability in thermophilic systems. Briefly, the metho...

BACKGROUND Thermostable enzymes have several benefits in lignocellulose processing. In particular, they potentially allow the use of increased substrate concentrations (because the substrate viscosity decreases as the temperature increases), resulting in improved product yields and reduced capital and processing costs. A short pre-hydrolysis step at an elevated temperature using thermostable en...

Using directed evolution and site-directed mutagenesis, we have isolated a highly thermostable variant of Aspergillus niger glucoamylase (GA), designated CR2-1. CR2-1 includes the previously described mutations Asn20Cys and Ala27Cys (forming a new disulfide bond), Ser30Pro, Thr62Ala, Ser119Pro, Gly137Ala, Thr290Ala, His391Tyr and Ser436Pro. In addition, CR2-1 includes several new putative therm...

Efficient enzymatic hydrolysis of lignocellulose to fermentable sugars requires a complete repertoire of biomass deconstruction enzymes. Hemicellulases play an important role in hydrolyzing hemicellulose component of lignocellulose to xylooligosaccharides and xylose. Thermostable xylanases have been a focus of attention as industrially important enzymes due to their long shelf life at high temp...

D-Hydantoinase is currently employed as a biocatalyst for the production of optically pure D-amino acids, which are intermediates for the synthesis of semisynthetic antibiotics, peptide hormone, pyrethroids, and pesticides. In the process developed by the Yamada group,1,2 DL-5-substituted hydantoin is asymmetrically hydrolyzed to the N-carbamoyl-D-amino acid by D-specific hydantoinase, and this...

A gene (tap) encoding a thermostable alkaline phosphatase from the thermophilic bacterium Thermus thermophilus XM was cloned and sequenced. It is 1506 bp long and encodes a protein of 501 amino acid residues with a calculated molecular mass of 54.7 kDa. Comparison of the deduced amino acid sequence with other alkaline phosphatases showed that the regions in the vicinity of the phosphorylation s...