Production of phytase by a marine yeast Kodamaea ohmeri BG3 in an oats medium: optimization by response surface methodology.

Statistical experimental designs were applied for the optimization of phytase production by a marine yeast Kodamaea ohmeri BG3 in a cost-effective oats medium. Using Plackett-Burman design, oats, ammonium sulfate and initial pH were identified as significant factors and these factors were subsequently optimized using a central composite design (CCD). The optimum variables that supported maximum enzyme activity were oats 1.0%, ammonium sulfate 2.3%, glucose 2.0%, NaCl 2.0% and initial pH 6.3. The validity of the optimized variables was verified in shake-flasks level. An overall 9-fold enhancement in phytase activity (62.0-->575.5 U/ml) was attained due to the optimization.

Bioethanol production from hydrolysates of inulin and the tuber meal of Jerusalem artichoke by Saccharomyces sp. W0.

It has been confirmed that Saccharomyces sp. W0 can produce high concentration of ethanol. However, this yeast strain cannot secrete inulinase. Therefore, in this study, inulin was hydrolyzed into reducing sugar by the recombinant inulinase produced by Pichia pastoris X-33/pPICZaA-INU1. It was found that 38.2U of the recombinant inulinase per gram of inulin was suitable for the inulin hydrolysis and ethanol production by Saccharomyces sp. W0 and the fermentation period was 120 h. At the end of the fermentation, over 14.6 ml of ethanol per 100ml of the fermented medium was produced, the ethanol productivity was over 0.384 g of ethanol/g of inulin and over 98.8% of total sugar was utilized. When the Saccharomyces sp. W0 was grown in the mixture of 4.0% hydrolysate of soybean meal and 20.0% of the hydrolysate of inulin for 120 h, over 14.9 ml of ethanol per 100ml of the fermented medium was yielded, the ethanol productivity was over 0.393 g of ethanol/g of inulin and 98.9% of total sugar was used by the yeast strain. When Saccharomyces sp. W0 carrying the same inulinase gene was grown in the medium containing 50 g of the tuber meal of Jerusalem artichoke per 100ml for 144 h, over 12.1+/-0.35%ml of ethanol per 100ml of the fermented medium was yielded, the ethanol productivity was 0.319+/-0.9 g of ethanol/g of sugar and 3.7% (w/v) of total sugar and 0.5% (w/v) of reducing sugar were left in the fermented media.

Siderophore production by the marine-derived Aureobasidium pullulans and its antimicrobial activity.

Over 300 yeast strains isolated from different marine environments were screened for their ability to produce siderophore. Among them, only the yeast strain HN6.2 which was identified to be Aureobasidium pullulans was found to produce high level of the siderophore. Under the optimal conditions, this yeast strain could produce 1.1mg/ml of the siderophore. The crude siderophore produced by the yeast strain HN6.2 was able to inhibit cell growth of Vibrio anguillarum and Vibrio parahaemolyticus, isolated from the diseased marine animals.

Trehalose synthesis in Saccharomycopsis fibuligera does not respond to stress treatments.

Synthesis of trehalose in Saccharomycopsis fibuligera sdu under various stress conditions was investigated. Neither the activation of trehalose-6-phosphate synthase (SfTPS1) nor the change in trehalose content was observed under stress exposure of S. fibuligera sdu cells. The results of reverse transcription polymerase chain reaction, which was performed with the specific primers designed to target the SfTPS1 gene fragment cloned from this strain, also showed that all stress treatments did not increase the expression of SfTPS1 gene. These results demonstrated that synthesis of trehalose in response to stress conditions in S. fibuligera sdu clearly differs from that of Saccharomyces cerevisiae and most other fungi. The phylogenetic analysis of the amino acid sequence deduced from the SfTPS1 gene fragment showed that the SfTPS1 sequence formed a separate family that was far related to S. cerevisiae TPS1. The yeast strain, which can accumulate a large amount of trehalose under normal growth conditions, has many applications and TPS1 gene in such strain may have unique use in transgenic organisms.

Trehalose accumulation in a high-trehalose-accumulating mutant of Saccharomycopsis fibuligera sdu does not respond to stress treatments.

The isolation of high-trehalose-accumulating mutant A11 from Saccharomycopsis fibuligera sdu has been previously described. In this paper, accumulation of trehalose under various stress conditions in S. fibuligera A11 was investigated. Neither activation of trehalose-6-phosphate synthase (SfTps1) nor change in trehalose content was observed under stress exposure of S. fibuligera A11 cells. A fragment of the Sftps1 gene in this strain was also cloned by degenerate PCR using the CoDeHOP strategy and multiply-aligned Tps1 sequences. This sequence allowed us to investigate the expression of the Sftps1 gene, which was also kept constant under the various stress conditions. Altogether, these results indicate that trehalose metabolism in S. fibuligera A11 in response to stress conditions clearly differs from that of Saccharomyces cerevisiae and most other fungi. The expression of the Sftps1 gene was not responsive to different stress treatments.