High-efficiency chromosomal integrative amplification strategy for overexpressing α-amylase in Bacillus licheniformis.

Bacillus licheniformis is a well-known platform strain for production of industrial enzymes. However, the development of genetically stable recombinant B. licheniformis for high-yield enzyme production is still laborious.

Exploitation of ammonia-inducible promoters for enzyme overexpression in Bacillus licheniformis.

Ammonium hydroxide is conventionally used as an alkaline reagent and cost-effective nitrogen source in enzyme manufacturing processes. However, few ammonia-inducible enzyme expression systems have been described thus far. In this study, genomic-wide transcriptional changes in Bacillus licheniformis CBBD302 cultivated in media supplemented with ammonia were analyzed, resulting in identification of 1443 differently expressed genes, of which 859 genes were upregulated and 584 downregulated.

Optimization of cultivation medium and cyclic fed-batch fermentation strategy for enhanced polyhydroxyalkanoate production by Bacillus thuringiensis using a glucose-rich hydrolyzate.

The accumulation of petrochemical plastic waste is detrimental to the environment. Polyhydroxyalkanoates (PHAs) are bacterial-derived polymers utilized for the production of bioplastics. PHA-plastics exhibit mechanical and thermal properties similar to conventional plastics.

A unique xylose reductase from Thermomyces lanuginosus: Effect of lignocellulosic substrates and inhibitors and applicability in lignocellulosic bioconversion.

In this study, the xylose reductase gene (XRTL) from Thermomyces lanuginosus SSBP was expressed in Pichia pastoris GS115 and Saccharomyces cerevisiae Y294. The purified 39.2 kDa monomeric enzyme was optimally active at pH 6.

Comparative biocontrol ability of chitinases from bacteria and recombinant chitinases from the thermophilic fungus Thermomyces lanuginosus.

Microbial chitinases (EC 3.2.1.

Concomitant production of chitosan and lipids from a newly isolated Mucor circinelloides ZSKP for biodiesel production.

A newly-isolated oleaginous fungus Mucor circinelloides ZSKP concurrently yielded 21.4% lipids and 11.2% chitosan per gram of biomass.

Simultaneous removal of heavy metals and cyanate in a wastewater sample using immobilized cyanate hydratase on magnetic-multiwall carbon nanotubes.

Global environmental problems allied with waste management require novel approaches for the simultaneous removal of heavy metals and other associated compounds including cyanate. In this study, iron-oxide filled multi-walled carbon nanotubes (m-MWCNTs) were successfully synthesized and characterized by field emission gun scanning electron microscopy (FEGSEM), high-resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD). The m-MWCNTs were amino-functionalized for the covalent immobilization of a recombinant cyanate hydratase (rTl-Cyn), and were characterized by fourier transform infrared (FTIR) spectroscopy.

Improved ethanol productivity from lignocellulosic hydrolysates by Escherichia coli with regulated glucose utilization.

Background: Lignocellulosic ethanol could offer a sustainable source to meet the increasing worldwide demand for fuel. However, efficient and simultaneous metabolism of all types of sugars in lignocellulosic hydrolysates by ethanol-producing strains is still a challenge.

Results: An engineered strain Escherichia coli B0013-2021HPA with regulated glucose utilization, which could use all monosaccharides in lignocellulosic hydrolysates except glucose for cell growth and glucose for ethanol production, was constructed.

A novel strategy for production of ethanol and recovery of xylose from simulated corncob hydrolysate.

Objectives: To develop a xylose-nonutilizing Escherichia coli strain for ethanol production and xylose recovery.

Results: Xylose-nonutilizing E. coli CICIM B0013-2012 was successfully constructed from E.

Exploring the Metabolomic Responses of to Temperature Stress by Gas Chromatography/Mass Spectrometry.

Owing to its high protein secretion capacity, simple nutritional requirements, and GRAS (generally regarded as safe) status, is widely used as a host for the industrial production of enzymes, antibiotics, and peptides. However, as compared with its close relative , little is known about the physiology and stress responses of . To explore its temperature-stress metabolome, strains ATCC 14580 and B186, with respective optimal growth temperatures of 42°C and 50°C, were cultured at 42°C, 50°C, and 60°C and their corresponding metabolic profiles were determined by gas chromatography/mass spectrometry and multivariate statistical analyses.

Production of gellan gum, an exopolysaccharide, from biodiesel-derived waste glycerol by spp.

In the present study, biodiesel-derived waste glycerol (WG) was used for the isolation and production of gellan, an exopolysaccharide, on media containing WG as the main carbon source. Two bacterial isolates showed gellan producing potential which were identified as (Accession No. GI:724472387) and (GI:724472388) by 16S rRNA gene sequencing.

A novel strategy for the efficient removal of toxic cyanate by the combinatorial use of recombinant enzymes immobilized on aminosilane modified magnetic nanoparticles.

Cyanase detoxifies cyanate by transforming it to ammonia and carbon dioxide in a bicarbonate-dependent reaction, however, dependence on bicarbonate limits its utilization in large-scale applications. A novel strategy was therefore developed for overcoming this bottleneck by the combined application of cyanase (rTl-Cyn) and carbonic anhydrase (rTl-CA). The synergistic effect of rTl-Cyn and rTl-CA could reduce the dependence of bicarbonate by 80%, compared to using rTl-Cyn alone.

Expression of a novel recombinant cyanate hydratase (rTl-Cyn) in Pichia pastoris, characteristics and applicability in the detoxification of cyanate.

A recombinant Pichia pastoris harbouring the cyanate hydratase gene (rTl-Cyn) from the thermophilic fungus Thermomyces lanuginosus SSBP yielded a high titre of extracellular cyanate hydratase (100±13UmL) which was ∼10-fold higher than the native fungal strain. The purified rTl-Cyn had a molecular mass of ∼20kDa on SDS-PAGE, with K, V, k and k/K values of 0.34mM, 2857.

Thermo-acid-stable phytase-mediated enhancement of bioethanol production using Colocasia esculenta.

Phytase production by the thermophilic mould Thermomyces lanuginosus SSBP was enhanced 8.56-fold in submerged fermentation, which was further improved in fed-batch cultivations. The protein was purified to homogeneity using ammonium sulphate precipitation, Resource Q anion exchange and Superdex gel-filtration chromatography, with an overall purification of 24.

Production, characteristics and applications of phytase from a rhizosphere isolated Enterobacter sp. ACSS.

Optimization of process parameters for phytase production by Enterobacter sp. ACSS led to a 4.6-fold improvement in submerged fermentation, which was enhanced further in fed-batch fermentation.

Chitinase from Thermomyces lanuginosus SSBP and its biotechnological applications.

Chitinases are ubiquitous class of extracellular enzymes, which have gained attention in the past few years due to their wide biotechnological applications. The effectiveness of conventional insecticides is increasingly compromised by the occurrence of resistance; thus, chitinase offers a potential alternative to the use of chemical fungicides. The thermostable enzymes from thermophilic microorganisms have numerous industrial, medical, environmental and biotechnological applications due to their high stability for temperature and pH.

Cloning, expression, and molecular dynamics simulations of a xylosidase obtained from Thermomyces lanuginosus.

The aim of this study was to clone, express, and characterize a β-xylosidase (Tlxyn1) from the thermophilic fungus Thermomyces lanuginosus SSBP in Pichia pastoris GS115 as well as analyze optimal activity and stability using computational and experimental methods. The enzyme was constitutively expressed using the GAP promoter and secreted into the medium due to the alpha-mating factor secretion signal present on the expression vector pBGPI. The 1276 bp gene consists of an open reading frame that does not contain introns.

Purification and Characterization of an Endoinulinase from pv. KM 24 Mutant.

An extracellular endoinulinase from pv. KM 24 mutant was purified to homogeneity by gel filtration chromatography and showed a specific activity of 119 U/mg. The optimum pH and temperature of the purified enzyme were found to be 6.

Secretome analysis of the thermophilic xylanase hyper-producer Thermomyces lanuginosus SSBP cultivated on corn cobs.

Thermomyces lanuginosus is a thermophilic fungus known for its ability to produce industrially important enzymes including large amounts of xylanase, the key enzyme in hemicellulose hydrolysis. The secretome of T. lanuginosus SSBP was profiled by shotgun proteomics to elucidate important enzymes involved in hemicellulose saccharification and to characterise the presence of other industrially interesting enzymes.

Creation of thermostable and alkaline stable xylanase variants by DNA shuffling.

Mutant xylanases, G41 and G53, were generated by random mutagenesis of Thermomyces lanuginosus xylanase DSM 5826 (xynA) in a previous study. Incubation at 90 min showed that G41 had 75% activity at 80 °C and G53 had 93% activity at pH 10. In order to create xylanase variants possessing both thermal and alkaline stability in a single enzyme, G41 and G53 served as templates for DNA shuffling using the StEP recombination method.

Biodegradation of glycerol using bacterial isolates from soil under aerobic conditions.

Glycerol, a non-biodegradable by-product during biodiesel production is a major concern to the emerging biodiesel industry. Many microbes in natural environments have the ability to utilize glycerol as a sole carbon and energy source. The focus of this study was to screen for microorganisms from soil, capable of glycerol utilization and its conversion to value added products such as ethanol and 1,3-propanediol (1,3-PDO).

Xylanase Superproducer: Genome Sequence of a Compost-Loving Thermophilic Fungus, Thermomyces lanuginosus Strain SSBP.

We report here the draft genome sequence of Thermomyces lanuginosus strain SSBP, which was isolated from soil in South Africa. This fungus produces the largest amount of xylanase ever reported in the literature.