Yeasts and yeast-like fungi associated with tree bark: diversity and identification of yeasts producing extracellular endoxylanases.

A total of 239 yeast strains was isolated from 52 tree bark samples of the Medaram and Srisailam forest areas of Andhra Pradesh, India. Based on analysis of D1/D2 domain sequence of 26S rRNA gene, 114 strains were identified as ascomycetous; 107 strains were identified as basidiomycetous yeasts; and 18 strains were identified as yeast-like fungi. Among the ascomycetous yeasts, 51% were identified as members of the genus Pichia, and the remaining 49% included species belonging to the genera Clavispora, Debaryomyces, Kluyveromyces, Hanseniaspora, Issatchenkia, Lodderomyces, Kodamaea, Metschnikowia, and Torulaspora. The predominant genera in the basidiomycetous yeasts were Cryptococcus (48.6%), Rhodotorula (29%), and Rhodosporidium (12.1%). The yeast-like fungi were represented by Aureobasidium pullulans (6.7%) and Lecythophora hoffmanii (0.8%). Of the 239 yeast strains tested for Xylanase, only five strains of Aureobasidium sp. produced xylanase on xylan-agar medium. Matrix-assisted laser desorption ionization-time of flight analysis and N-terminal amino-acid sequence of the xylanase of isolate YS67 showed high similarity with endo-1-4-beta-xylanase (EC 3.2.1.8) of Aureobasidium pullulans var. melanigenum.

Modelling and optimization of fermentation factors for enhancement of alkaline protease production by isolated Bacillus circulans using feed-forward neural network and genetic algorithm.

AIM: Modelling and optimization of fermentation factors and evaluation for enhanced alkaline protease production by Bacillus circulans. METHODS AND RESULTS: A hybrid system of feed-forward neural network (FFNN) and genetic algorithm (GA) was used to optimize the fermentation conditions to enhance the alkaline protease production by B. circulans. Different microbial metabolism regulating fermentation factors (incubation temperature, medium pH, inoculum level, medium volume, carbon and nitrogen sources) were used to construct a '6-13-1' topology of the FFNN for identifying the nonlinear relationship between fermentation factors and enzyme yield. FFNN predicted values were further optimized for alkaline protease production using GA. The overall mean absolute predictive error and the mean square errors were observed to be 0.0048, 27.9, 0.001128 and 22.45 U ml(-1) for training and testing, respectively. The goodness of the neural network prediction (coefficient of R(2)) was found to be 0.9993. CONCLUSIONS: Four different optimum fermentation conditions revealed maximum enzyme production out of 500 simulated data. Concentration-dependent carbon and nitrogen sources, showed major impact on bacterial metabolism mediated alkaline protease production. Improved enzyme yield could be achieved by this microbial strain in wide nutrient concentration range and each selected factor concentration depends on rest of the factors concentration. The usage of FFNN-GA hybrid methodology has resulted in a significant improvement (>2.5-fold) in the alkaline protease yield. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study helps to optimize enzyme production and its regulation pattern by combinatorial influence of different fermentation factors. Further, the information obtained in this study signifies its importance during scale-up studies.

Isolation and characterization of xylitol-producing yeasts from the gut of colleopteran insects.

A total of 35 yeasts were isolated from the gut of beetles collected from Hyderabad city, India. Twenty of these yeasts utilized xylose as a sole carbon source but only 12 of these converted xylose to xylitol. The ability to convert xylose to xylitol varied among the isolates and ranged from 0.12 to 0.58 g/g xylose. Based on the phenotypic characteristics and phylogenetic analysis of the D1/D2 domain sequence of 26S rRNA gene, these isolates were identified as members of Pichia, Candida, Issatchenkia, and Clavispora. Strain YS 54 (CBS 10446), which was phylogenetically similar to Pichia caribbica and which formed hat-shaped ascospore characteristics of the genus Pichia, was the best xylitol producer (0.58 g xylitol/g xylose). YS 54 was also capable of producing xylitol from sugarcane bagasse hydrolysate and the efficiency of conversion was 0.32 g xylitol/g xylose after 20 cycles of adaptation in medium containing sugarcane bagasse hydrolysate.

Microbacterium indicum sp. nov., isolated from a deep-sea sediment sample from the Chagos Trench, Indian Ocean.

Two bacterial strains, BBH6(T) and BBH9, were isolated from a deep-sea sediment sample collected from the Chagos Trench, Indian Ocean, at a depth of 5904 m. The two strains were closely related in their 16S rRNA gene sequences (99.7 %), belonged to one genomic species and were virtually identical at the phenotypic level. Microbacterium barkeri DSM 20145(T) was the nearest phylogenetic neighbour to the new isolates, with 16S rRNA gene sequence similarity levels of 97.2-97.4 %. The new isolates exhibited levels of DNA-DNA relatedness of 32-34 % to M. barkeri and differed from it in a number of phenotypic characteristics. Therefore, it is suggested that strains BBH6(T) and BBH9 represent a novel species of the genus Microbacterium, for which the name Microbacterium indicum sp. nov. is proposed. The type strain is BBH6(T) (=LMG 23459(T)=IAM 15355(T)).

L-asparaginase production by isolated Staphylococcus sp. - 6A: design of experiment considering interaction effect for process parameter optimization.

AIMS: Evaluation of fermentation process parameter interactions for the production of l-asparaginase by isolated Staphylococcus sp. - 6A. METHODS AND RESULTS: Fractional factorial design of experimentation (L18 orthogonal array of Taguchi methodology) was adopted to optimize nutritional (carbon and nitrogen sources), physiological (incubation temperature, medium pH, aeration and agitation) and microbial (inoculum level) fermentation factors. The experimental results and software predicted enzyme production values were comparable. CONCLUSION: Incubation temperature, inoculum level and medium pH, among all fermentation factors, were major influential parameters at their individual level, and contributed to more than 60% of total l-asparaginase production. Interaction data of selected fermentation parameters could be classified as least and most significant at individual and interactive levels. Aeration and agitation were most significant at interactive level, but least significant at individual level, and showed maximum severity index and vice versa at enzyme production. SIGNIFICANCE AND IMPACT OF THE STUDY: All selected factors showed impact on l-asparaginase enzyme production by this isolated microbial strain either at the individual or interactive level. Incubation temperature, inoculum concentration, pH of the medium and nutritional source (glucose and ammonium chloride) had impact at individual level, while aeration, agitation and incubation time showed influence at interactive level. Significant improvement (ninefold increase) in enzyme production by this microbial isolate was noted under optimized environment.

Xylitol production from corn fiber and sugarcane bagasse hydrolysates by Candida tropicalis.

A natural isolate, Candida tropicalis was tested for xylitol production from corn fiber and sugarcane bagasse hydrolysates. Fermentation of corn fiber and sugarcane bagasse hydrolysate showed xylose uptake and xylitol production, though these were very low, even after hydrolysate neutralization and treatments with activated charcoal and ion exchange resins. Initial xylitol production was found to be 0.43 g/g and 0.45 g/g of xylose utilised with corn fiber and sugarcane bagasse hydrolysate respectively. One of the critical factors for low xylitol production was the presence of inhibitors in these hydrolysates. To simulate influence of hemicellulosic sugar composition on xylitol yield, three different combinations of mixed sugar control experiments, without the presence of any inhibitors, have been performed and the strain produced 0.63 g/g, 0.68 g/g and 0.72 g/g of xylose respectively. To improve yeast growth and xylitol production with these hydrolysates, which contain inhibitors, the cells were adapted by sub culturing in the hydrolysate containing medium for 25 cycles. After adaptation the organism produced more xylitol 0.58 g/g and 0.65 g/g of xylose with corn fiber hydrolysate and sugarcane bagasse hydrolysate respectively.

Effect of hydrocarbons and other parameters on hydrocarbon-utilizing Pichia angusta MTCC-225.

Pichia angusta MTCC-225, a catalase-positive yeast that utilizes methanol and lighter hydrocarbons, is the subject of this investigation. An orthogonal experimental design L16 was used to investigate the effects of methanol, a gas mixture, zero air, temperature, agitation, and salts solution on hydrocarbon utilizing P. angusta. QUALITEK-4 Software was used for automatic design and analysis of the experimental results. Among the various parameters tested, agitation contributed the highest influence (56.5%). Zero air, methanol concentration, and gas mixture showed a moderate influence on the growth of P. angusta. Methanol concentration and gas mixture showed a 10.91 and 10.12% influence, respectively, on yeast growth. Zero air played an important role, with a 15.19% influence on the utilization of hydrocarbon.

Optimization of alkaline protease production by Bacillus sp. using Taguchi methodology.

Optimization of alkaline protease production parameters by Bacillus sp. was investigated using Taguchi methodology. The pH of the medium was observed to be the most significant factor among all selected optimization parameters at an individual level. The combinatorial influence of least significant factors, inoculum level and salt solution concentration (at the individual level), resulted in an interacting severity index of 76%, suggesting their interactive role in the regulation of protease production in this microbial species. Protease production could be improved more than 100% with Taguchi's optimized conditions of the medium composition by this microorganism.