• KSBB Journal
• /
• v.21 no.5
• /
• pp.376-383
• /
• 2006

Pine needle(Pinus densiflora sieb, et zucc) extract has been used to improve cardiovascular disorders, detoxification of nicotine, the infirmities of age and curing diseases of unidentified symptoms. It has various useful components including amino acids, vitamin C, terpenoids and chlorophyll. In this study we have identified 8 different yeast strains that are developed spontaneously causing self fermentation in the extract. The self-fermented pine extract(SFPE) inhibited the growth of some bacterial strains like E. coli, Bacillus subtilis and Staphylococcus aureus. The SFPE($0.2{\mu}{\ell}/ml{\sim}0.3{\mu}{\ell}/ml$) showed 90% NBT superoxide scavenging activities which is similar for all tested samples of different ages. The 7 years old SFPE(0.15 mg/ml and 0.3 mg/ml) caused relaxation of spontaneous contraction and relaxation rhythm of thoracic arterial tissues from rat. Therefore, SFPE has useful effects such as antibacterial, antioxidant and improved blood circulation and could be a good source of functional food development.

• BMB Reports
• /
• v.37 no.3
• /
• pp.282-291
• /
• 2004

Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.10
• /
• pp.1012-1020
• /
• 2011

A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.

• Journal of the Korean Wood Science and Technology
• /
• v.39 no.1
• /
• pp.75-85
• /
• 2011

The main purpose of this study is to evaluate the potential of producing bioethanol from yellow poplar ($Liriodendron$ $tulipifera$) wood chips by oxalic acid pretreatment and to examine the pretreatment conditions by response surface methodology (RSM). Based on $2^3$ factorial design, adjusted variables were reaction temperature ($^{\circ}C$), residence time (min), and acid loading (g/g), and a series of distinct 15 experimental conditions was organized with duplication at central point (total 16 performances). After pretreatment, simultaneous saccharification and fermentation (SSF) was subjected on solid fraction with yeast strain $Pichia$ $stipitis$. Maximum ethanol yields of the most samples were measured at 72 hours and applied to RSM as a dependent variable. 9.7 g/${\ell}$ of ethanol was produced from the solid pretreated at $180^{\circ}C$ for 40 min with 0.013 g/g of oxalic acid loading. According to the response surface methodology, it was determined that the temperature is the most governing factor via statistic analysis.

• Korean Journal of Microbiology
• /
• v.37 no.2
• /
• pp.170-175
• /
• 2001

In order to produce xylitol with high yield, experiments were carried out to develope xylitol dehydrogenase(XDH) defective mutant from P. stipitis and to investigate the xylitol fermentation characteristics of mutant strain. After treatment of P. stipitis with EMS, mutant PXM-4 was selected based on te XDH activity and xylitol production capability. Among the tested cosubstrates, galactose was selected as an adequate cosub-strate on xylitol production of mutant PXM-4. But with the increase in the concentration of galactose in the medium, xylitol production was decreased because the transport of xylose into cell was inhibited by galactose. The optimal concentration of galactose for the production of xylitol using 20 g/ι xylose was 20 g/ι Under this condition, maximum concentration of xylitol and yield were 14.4 g/ι and 97%, respectively. In order to prevent the inhibitory effect of xylose transport by galactose, galactose was fed with low concentration and the concentration of xylitol produced was increased up to 25 g/ι.

• Journal of Food Hygiene and Safety
• /
• v.22 no.2
• /
• pp.99-104
• /
• 2007

To defend putrifaction of the processed rice cake from gas-forming yeast during storage and distribution it needed to reduce and remove them. The sanitizers of ethanol and organic acids were applied on Pichia anomala, Candida tropicalis, and isolated yeasts from the putrified cut rice cake. Although growth inhibition effect by the sanitizer of 20% ethanol, 1% acetic acid, or 1% lactic acid respectively were very low, the combined sanitizer of 20% ethanol and 1% acetic acid, or 1% lactic acid showed very high sterilizing effect toward the yeasts. Six log cfu/ml of the yeast was reduced with this combined sanitizers for 30 minutes. In addition, the combined sanitizer heated from 20 to $50^{\circ}C$ had more the increased sterility. Therefore, the sanitizer of the combined ethanol with the acetic acid or the lactic acid for 30 minutes at $50^{\circ}C$ might reduce or sterilize the putrifying yeast at the processed rice cake. The result might be also applied to the effective pre-treatment of many agricultural food stuffs, against yeast, especially unsterilized stuffs, without any hazards from the special sanitizers and nutritional loss from harsh sterilization.

• KSBB Journal
• /
• v.20 no.2 s.91
• /
• pp.93-99
• /
• 2005

Polyunsaturated fatty acids, that is PUFAs, are important constituents of membranes particularly found in the retina and central nervous system. In microorganism-based PUFAs biosynthesis, the genus Thraustochytrids is well evaluated for their potential as a promising candidate in the practical production of PUFAs, such as AA and DHA. In this study, we attempted to optimize a method of total nucleic acid extraction from this microorganism as a preliminary experiment. Using the extracted nucleic acid and degenerated primers for direct PCR, we isolated a $\Delta5$ desaturase gene that contained 1320-nucleotide and encoded 439 amino acids. This gene exhibited an expected function, when expressed in P. pastoris in the presence of appropriate exogenous substrate, as an evidence for $\Delta5$ desaturase activity (conversion of DGLA to AA). These results and information could provide a basis for the construction of engineered strains suitable for the practical production of PUFAs.

• The Korean Journal of Mycology
• /
• v.44 no.3
• /
• pp.138-144
• /
• 2016

Twelve unrecorded yeasts, Pseudozyma prolifica HL9-1, Trichosporon coremiiforme NS19-2, Candida cretensis SA4-1, Cryptococcus diffluens TJ4-3, Cryptococcus pinus YB17-2, Candida vartiovaarae DD2-5, Pichia galeiformis DM3-5, Candida pseudolambica JW2-3, Trichosporon xylopini NS5-1, Trichosporon moniliiforme NS5-7, Tetrapisispora iriomotensis NS14-2, and Tetrapisispora nanseiensis SA17-1, were screened among 97 yeasts from soils of Chungcheongnam-do and Daejeon metropolitan city, Korea. These yeasts were oval or ellipsoidal and had a budding system for vegetative reproduction. They grew well in yeast extract-peptone-dextrose (YPD) medium and, in particular, Tetrapisispora iriomotensis NS14-2 and Candida cretensis SA4-1 grew well in 10% NaCl-containing YPD broth. Nine strains, including Trichosporon coremiiforme NS19-2, assimilated xylose and four yeast strains, such as Candida vartiovaarae DD2-5, also assimilated lactose. Physiological functionalities of cell-free extracts and supernatants from two halophilic unrecorded yeasts, Candida cretensis SA4-1 and Tetrapisispora iriomotensis NS14-2, were investigated. Cell-free extracts from Candida cretensis SA4-1 and Tetrapisispora iriomotensis NS14-2 exhibited 71.3% and 68.4% antihypertensive angiotensin I-converting enzyme inhibitory activity.

• Food Engineering Progress
• /
• v.14 no.4
• /
• pp.292-298
• /
• 2010

Wheat germ contains the glycosylated forms of 2-methoxy-p-benzoquinone (2-MBQ) and 2,6-dimethoxy-p-benzoquinone (2,6-DMBQ), both of which have antimicrobial and immunostimulatory effects. Conversion of glycosylated 2-MBQ and 2,6-DMBQ to their more functional unglycosylated forms requires enzymatic action of $\beta$-glucosidase. We investigated the applications of lactic acid bacteria and yeast that produce $\beta$-glucosidase as starters for production of unglycosylated 2-MBQ and 2,6-DMBQ from wheat germ. Lactobacillus zeae and Pichia pijperi were selected through $\beta$-glucosidase enzyme assays for 37 yeast strains and five strains of lactic acid bacteria. Lb. zeae was more efficient than P. pijperi at producing 2-MBQ and 2,6-DMBQ from wheat germ. After 48 hr of fermentation with a mixed culture of Lb. zeae and P. pijperi, the concentration of 2-MBQ was 0.46${\pm}$0.07 mg/g, indicating an approximately 1.6-fold higher concentration than that obtained by pure culture of Lb. zeae. However, the concentration of 2,6-DMBQ was not significantly enhanced by fermentation with a mixed culture of Lb. zeae and P. pijperi.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.4
• /
• pp.564-569
• /
• 2001

This study was carried out in order to investigate the characteristics of xylitol fermentation by a xylitol dehydrogenase defective mutant PXM-4 of P stipitis CBS 5776 and to determime optimum conditions for the high yield ofxylitol production from xylose. Gluconic acid was selected as a co substrate for the xylitol fermentation, since gluconic acid neither blocked xylose transport nor repressed xylose reductase expression. An increase of gluconic acid concentration reduced the rates of xylitol production and cell growth by decreasing medium pH, and the optimal concentration of gluconic acid was determined to be 20 gll with approximately 100% xylitol conversion yield. A fed-batch cell culture resulted in a 44.8 g/l xylitol concentration with 100% yield, based on the amount of xylose consumed.