• Journal of the Korean Applied Science and Technology
• /
• v.27 no.2
• /
• pp.114-122
• /
• 2010

Hydroxy fatty acids have gained important attentions because of their special properties such as higher viscosity and reactivity compared to non-hydroxy fatty acids. The new bacterial isolate Pseudomonas aeruginosa (PR3) had been well studied to produce mono-, di-, and tri-hydroxy fatty acids from different unsaturated fatty acids. Of those hydroxy fatty acids, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was produced with high yield from oleic acid by P. aeruginosa PR3. Mostly, the substrates used for the production of hydroxy fatty acid by microorganisms were free fatty acids or synthetic lipids. However, it is highly plausible to use vegetable oils containing oleic acid such as olive oil for DOD production by P. aeruginosa PR3. As a first step to address the utilization of olive oil as a substrate for DOD production, we tried to determine the effect of vessel type on DOD production from olive oil by P. aeruginosa PR3. Of two different flask types of normal flask and baffled-flask, baffled-flask was highly effective for DOD production with all the media tested. Maximum increase of productivity by baffled-flask represented 221% with the medium supplemented with whey powder instead of lactose. Results from this study demonstrated that vessel type and medium composition could be both significant factors for DOD production from olive oil by P. aeruginosa PR3.

• KSBB Journal
• /
• v.15 no.6
• /
• pp.566-572
• /
• 2000

Experimental studies were carried out to develop an osmotolerant mutant of Candida magnoliae JH and to determine the optimum concentrations of carbon and nitrogen sources to improve erythritol yield and productivity. Mutants of C. magnoliae JH were prepared by treatment with ethylmethane sulfonate, and osmotolerant mutants were isolated from the agar plate colonies containing 2.5 M KCI. Among the mutants isolated, one mutant M26 was finally selected based on erythritol yield and productivity. In shake flask culture, glucose proved to be the best carbon source and the optimum yeast extract concentration was 5 g/L based on 100 g/L glucose. The erythritol yield and productivity of mutant M26 were greater than wild type in 100 g/L glucose medium. in the fermentation experiments, erythritol production increased with increased glucose concentration, up to a limit of 250 g/L. The maximum concentration of erythritol achieved 127.5 g/L, and the yield and productivity of erythritol production were 51.0% and 0.63 g/L-h, respectively.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.5
• /
• pp.722-728
• /
• 2002

Eight strains producing bacterial cellulose (BC) were isolated from rotten fruits and traditionally fermented vinegars. One of the isolated strains from the rotten grape in Gwangju, Korea, maintained a relatively stable BC production in shaking cultures. This isolated strain proved to be Acetobacter xylinum, based on several biochemical and morphological tests. It was shown that the slant-baffled flask was more efficient than the conventional flask for the BC production in shaking cultures. To determine the most suitable carbon and nitrogen sources for the production of BC, various compounds were examined. Fructose was found to be the most effective carbon source with an optimal concentration of 2%. Mixed carbon source (glucose:fructose=1:3) was also better than glucose or fructose alone. Optimal nitrogen source, when basal medium was used, was 10% (v/v) com steep liquor (CSL). When com steep liquor was used with a mixed carbon source (glucose:fructose=1 :3),4% CSL exhibited the best BC production. Based on these results, a defined medium was developed for the BC production by Acetobacter xylinum KJ-1. When this medium was used under optimal culture conditions, the BC production was 7.2 g/1, which was approximately 3 times higher than that with the traditional HS medium.

• Microbiology and Biotechnology Letters
• /
• v.38 no.1
• /
• pp.40-45
• /
• 2010

The gene encoding $\beta$-agarase (agaB) which hydrolyzes $\beta$-1,4 linkages of agarose from Zobellia galactanivorans was cloned and fused to Saccharomyces cerevisiae mating factor alpha-1 secretion signal ($MF{\alpha}1$), in which the transcription of $MF{\alpha}1$-AgaB was under the control of AOX1 (alcohol oxidase 1, methanol inducible) promoter. The constructed plasmid pPIC-AgaB (9 kb) was integrated into HIS4 gene locus of Pichia pastoris genome. Successful integration was confirmed by performing colony PCR. The transformed cells showed red halos around its colonies in methanol agar plate by adding iodine solution, indicating the active expression of agaB in P.pastoris. By SDS-PAGE and zymographic analysis, the molecular weight of $\beta$-agarase was estimated to be a 53 kDa and about 15% N-linked glycosylation was occurred. The activity of extracellular $\beta$-agarase reached 1.34, 1.42 and 1.53 units/mL by inducing 0.1, 0.5, and 1% methanol, respectively, at baffled flask culture of P.pastoris GS115/pPIC-AgaB for 48 hr. Most of the enzyme activity was found in the extacellular fraction and the secretion efficiency showed 98%. Thermostability of recombinant $\beta$-agarase was also increased by glycosylation.

• Journal of Life Science
• /
• v.23 no.7
• /
• pp.863-868
• /
• 2013

The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and $MF{\alpha}_{opt}$ s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the $MF{\alpha}_{opt}$ s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.1
• /
• pp.69-74
• /
• 2000

The medium for erythritol production by Torula sp. in a 500-ml baffled flask was optimized to be 300 g/I sucrose, 10 g/I yeast extract, 3 g/I $KH_2PO_4$, and 10 mg/I $CuSO_4{\cdot}5H_2O{\;}at{\;}34^{\circ}C$ with initial pH of 5.5. Using this optimal medium, erythritol of 166 g/I was obtained after 140 h of cultivation, corresponding to 55.3% of the erythritol yield from sucrose with a productivity of 1.11 g/I/h. Optimal concentrations of carbbon and nitrogen sources in a fermentor were higher than that in a flask due to the higher oxygen supply of the fermentor. Employing the medium containing 300 g/I or 400 g/I sucrose for the determination of optimal C/N ratio, the C/N ratio was found to be more important than the nitrogen concentration for effective erythritol production, The optimal ratio of yeast extract to sucrose (g/g) was 20. The yield and productivity of erythritol were maximal in the medium containing 400 g/I sucrose and 20 g/I yeast extract. when dissolved oxygen in the culture was increased, the cell mass increased but the erythritol production was manimal in the range of 5 to 10% of dissolved oxygen. Under the optimal the rane of 5 to 10% of dissolved oxygen. Under the optimal culture condition of the fermentor, a final erythritol concentration of 200 gI was obtained after 120 h with a yield of 50% and the productivity was 1.67 g/I/h. The yield was the highest among erythritol-producting microorganisms

• Journal of Life Science
• /
• v.26 no.12
• /
• pp.1376-1382
• /
• 2016

Xylitol is widely used in the food and medical industry. It is produced by the reduction of xylose (lignocellulosic biomass) in the Saccharomyces cerevisiae strain, which is considered genetically safe. In this study, the expression system of the GRE3 (YHR104W) gene that encodes xylose reductase was constructed to efficiently produce xylitol in the S. cerevisiae strain, and the secretory production of xylose reductase was investigated. To select a suitable promoter for the expression of the GRE3 gene, pGMF-GRE3 and pAMF-GRE3 plasmid with GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter for secretory production. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$, and $SEY2102{\Delta}trp1$/pGMF- GRE3 and $SEY2102{\Delta}trp1$/pAMF-GRE3 transformants were selected. In the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, the total activity of xylose reductase reached 0.34 unit/mg-protein when NADPH was used as a cofactor; this activity was 1.5 fold higher than that in $SEY2102{\Delta}trp1$/pAMF-GRE3 with ADH1 as the promoter. The secretion efficiency was 91% in both strains, indicating that most of the recombinant xylose reductase was efficiently secreted in the extracellular fraction. In a baffled flask culture of the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, 12.1 g/l of xylitol was produced from 20 g/l of xylose, and ~83% of the consumed xylose was reduced to xylitol.

• Microbiology and Biotechnology Letters
• /
• v.30 no.1
• /
• pp.68-72
• /
• 2002

To obtain a yeast mutant with high RNA content and high growth rate, Saccharomyces cerevisiae MTY62 was mutated with ethylmethane sulfonate. Among the selected mutants that were sensitive to the high concentration of KCl, M40-10 strain was finally selected due to its rapid cell growth and high RNA content in the tube and baffled-flask cultures. In the batch culture of M40-10 mutant, the maximum specific growth rate ($\mu_{max}$) of $0.38 h^{-1}$ , RNA concentration of 3210 mg-RNA/1, and RNA content of 183 mg-RNA/g-DCW were obtained, which were 23%, 15%, and 12% increased levels, respectively, compared to those of MTY62 parent strain. The intermittent fed-batch culture of M40-10 strain resulted in the maximum cell concentration of 35.6 g-DCW/1, RNA concentration of 5677 mg/1, and RNA content of 160 mg-RNA/g-DCW. Through the constant fed-batch culture, the maximum cell concentration of 46.4 g-DCW/1, RNA concentration of 6270 mg-RNA/1, and RNA content of 135 mg-RNA/g-DCW were obtained. At the 20 h culture time in the fed-batch cultures of M40-10 strain, the cell and RNA concentrations were increased by 30% and 10%, respectively, over the parent strain MTY62. In addition, it was also found that the accumulated RNA within the mutant cell was not degraded until the end of fed-batch cultivation, indicating that the M40-10 cell is a mutant with weak acidic RNase activity.y.

• Current Research on Agriculture and Life Sciences
• /
• v.14
• /
• pp.101-110
• /
• 1996

Using response surface methodology(RSM), the various conditions(agitation speed, air flow, glucose concentration) in jar fermentor culture were investigated to find the optimum conditions for maximum enzyme production. Central-composite-design was used to control the variable constant in the experiment. The glucose isomerase production of Steptomyces chibaensis J-59 was mostly affected by the air flow rate and glucose concentration. The estimated optimum conditions were as follows: 1% birchwood xylan, 1.5% CSL, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, pH 7.0; air flow, 2.2vvm; agitation speed, 587rpm; glucose concentration, 0.586%. Experimental values(7.43GIU/ml) for the enzyme production obtained from the given optimum conditions had a almost resemblane to response values(7.67GIU/ml) predicted by the RSM. The jar fermentor culture by the RSM produced xylose isomerase about 2.7 times as much as the baffled flask culture.