• KSBB Journal
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• v.26 no.6
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• pp.557-560
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• 2011

It has been known that Δ-desaturase (TAD5) in the biosynthetic pathway of long chain polyunsaturated fatty acids of Thraustochytrium aureumis responsible for the conversion of di-homo-${\gamma}$-linolenic acid (C20：4) into arachidonic acid (C20：4). The genetic sequence analysis on TAD5 of Thraustochytrium aureum ATCC34304 used in this study showed that it has two amino acid changes when compared to that of Thraustochytrium aureum TAD5 first reported in 2003. Accordingly, Thraustochytrium aureum ATCC34304 TAD5 was named TAD5_1. TAD5_1-inserted methylotropic Pichia pastoris was prepared and then cultured with a precursor fatty acid, di-homo-${\gamma}$-linolenic acid. GC analysis confirmed that a certain amount of the precursor fatty acid was converted into arachidonic acid. In this study, not only a recombinant Pichia pastoris with the typical activity of ${\Delta}5$-desaturase which plays an essential role in the biosynthesis of LCPUFAs was successfully made but also the preparationpotential of a recombinant Pichia pastoris strain which may synthesize eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) that are important in maintaining and improving human's brain function was proposed.

• Food Science and Biotechnology
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• v.16 no.2
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• pp.193-197
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• 2007

In order to investigate the effects of Pichia farinosa SKM -1, Pichia anomala SKM-T, Galactomyces geotrichum SJM-59, and Saccharomyces cerevisiae on extending the shelf life of kkakdugi, 4 kinds of lyophilized yeasts adding kkakdugis were prepared and stored at $4^{\circ}C$ for 30 days. Except S. cerevisiae adding group, 3 kinds of yeast adding groups were maintained their desirable levels (ca. pH 4.2 and 0.6% acidity) during the fermentation. The hardness of yeast adding groups was higher than those of control during the experiments. The number of yeast and the ratio of lactic acid against to total bacteria in P. farinosa SKM-1, P. anomala SKM-T, and G geotrichum SJM-59 adding groups were lower than that of control and/or S. cerevisiae adding group. Based on acidity, kkakdugi made with P. farinosa SKM-1, P. anomala SKM-T, and G geotrichum SJM-59 remained edible about 10 days longer than the control product.

• Journal of Life Science
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• v.12 no.4
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• pp.496-503
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• 2002

To develop a yeast strain of Pichia pastoris producing an antibacterial peptide, we have attempted the expression and secretion of an insect defensin. The nucleotide sequences corresponding to mature defensin were chemically synthesized by 6 oligomers, assembled in vitro and the synthesized gene was identified by nucleotide sequencing. The prepro sequence of yeast mating factor $\alpha$1 and the defensin gene were recombined into a Pichia expression vector, pPIC9K. The resulting plasmid, pPIDE, was transformed into P. pastoris GSl15 and transformants selected on histidine-deficient minimal plates were tested for antibacterial activity against Micrococcus luteus. Four strains with different antibacterial activity were selected for further analysis. Southern hybridization and RT-PCR verified the defensin gene was maintained and transcribed in a host. Four strains were cultivated in YPD broth for 96 hours to compare cell growth and antibacterial activity, They showed no difference in cell growth, however, each strain showed different antibacterial activity pattern with culture time. The maximal activity was about 550 AU/ $m\ell$.

• The Korean Journal of Mycology
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• v.41 no.4
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• pp.218-224
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• 2013

Of 1,100 yeast strains which were isolated from various Korean fermented foods, screened for phosphate solubilization, five strains showed the ability to solubilize tricalcium phosphate. The 26S rDNA domain D1-D2 sequence analysis revealed the identification of strain Y393 and Y524 as Pichia anomala (99.8 and 100% identity, respectively), Y669 as Pichia farinosa (100% identity), Y901 as Candida versatilis (100% identity), and Y1101 as Pichia subpelliculosa (100% identity). All the phosphate solubilizing strains showed mesophilic characteristics. The temperature range for growth of 4 strains was $20{\sim}35^{\circ}C$ and P. farinosa Y669 was able to grow up to $45^{\circ}C$. The strain C. versatilis Y907 was able to grow at pH range of 5.0~6.0 and showed halophilic characteristics with tolerance to 15% of NaCl concentration. The Phosphate solubilizing yeast strains were survived well in bed soil for 8 weeks which were maintained densities of $10^7{\sim}10^8$ cfu/g. The highest phosphate solubilizing activity was observed in P. subpelliculosa Y1101. It solubilized 697.2 ug/mL of phosphorus from tricalcium phosphate with decrease in pH from 6.8 to 4.37 after 11 days of inoculation.

• KSBB Journal
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• v.8 no.5
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• pp.452-456
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• 1993

This study was carried out to optimize the fermentation conditions for direct alcohol fermentation of xylose by Pichia stipitis CBS 5776. The best cell growth and the ethanol production were obtained under 0.05 VVM aeration and 300rpm agitation at $30^{\circ}C$ using 100 g/l xylose medium of the initial pH 5.0. In the above condition, the maximum specific growth rate and maximum cell concentration were 0.14hr-1 and $1.3 \times109$ cells/ml, respectively. Pichia stipitis CBS 5776 also produced 40.2g/l ethanol utilizing about 96% of 100g/l xylose after 72hr fermentation. At this point, the overall volumetric ethanol productivity was 0.56g/1-hr and the ethanol yield was 0.42 g-ethanol/g-xylose consumed, which corresponds to 82% of the theoretical yield.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.269-273
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• 2005

Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDl-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

• Korean Chemical Engineering Research
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• v.46 no.4
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• pp.764-768
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• 2008

Bovine testicular hyaluronidase PH-20 was cloned into pPIC9 vector and expressed in Pichia pastoris. Recombinant PH-20 was 75 kDa MW and 7460 units/L activity. Extracellular hyaluronidase activity was two times higher than that of intracellular activity. Non-buffered medium and $30^{\circ}C$ cultivation was favorable for PH-20 production. 1M sorbitol as an osmotic pressure and 0.3% methanol inducer increased cell growth and enzyme activity. 0.4 M arginine augmentation decreased the proteolytic degradation of recombinant hyaluronidase.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.275-287
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• 2000

Fermentation strategies for recombinant protein production in Pichia pastoris have been investigated and are reviewed here. Characteristics of the expression system, such as phenotypes and carbon utilization, are summarized. Recently reported results such as growth model establishment, app58lication of a methanol sensor, optimization of substrate feeding strategy, DOstat controller design, mixed feed technology, and perfusion and continuous culture are discussed in detail.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.171-174
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• 2005

The Pichia pastoris expression system is widely used for e production of recombinant proteins. In this study, we investigated the characteristics of cell growth and PLC production of recombinant Pichia pastoris. Especially, severed carbon sources like glycerol, glucose, sucrose, mannose, dextrose, xylitol, lactic acid, and acetic acid etc. were used to fermentations. The PLC activity was measured photometically.

• KSBB Journal
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• v.4 no.1
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• pp.57-61
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• 1989

The feasibility of separating Pichia stipitis from a fermentation broth using a hollow fiber membrane was evaluated. The permeate flux was affected by such parameters as cell concentration, pH, content of antifoam agents, suction pressure, and recirculation rate. A minor effect of temperature on the flux loss was also observed. A microcomputer-aided backflush was proven effective in alleviating membrane fouling and allowing long term separation of P. stipitis from a fermentation broth.