• Mycobiology
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• v.38 no.2
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• pp.153-155
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• 2010

To produce ginsenoside-$Rg_3$ enriched yeast from ginseng-steaming effluent (GSE), Lipomyces starkeyi, which tends to grow well in GSE, was cultured in sterilized GSE and its growth and production of ginsenoside-$Rg_3$ were determined. Growth of L. starkeyi was 86.1 mg per g GSE and its ginsenoside-$Rg_3$ contents was 0.013 mg per g GSE.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.665-683
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• 2002

A novel glucanhydrolase from a mutant of Lipomyces starkeyi KSM 22 has additional amylase activity besides mutanolytic activity and has been suggested as promising anti-plaque agent. It has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 glucanhydrolase are desirable for its application as a dental plaque control agent. In human experimental gingivitis　model and 6 month clinical trial, mouthrinsing with Lipomyces　starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effect was negligible. This study was aimed to evaluate the cytotoxic effect of Lipomyces starkeyi KSM 22 glucanhydrolase on human gingival fibroblasts. Primary culture of human gingival fibroblasts at the 4th to 6th passages were used. Glucanhydrolase solution was made from lyophilized glucanhydrolase powder from a mutant of Lipomyces stakeyi KSM 22 solved in PBS and added to DMEM medium to the final concentration of 0.5, 1, and 2 unit. Cells were exposed to glucanhydrolase solution or 0.1 % chlorhexidine and the cells cultured in DMEM with 10% FBS and 1% antibiotics as control. After exposure, the morphological change, cell attachment, and cell activity by MTT assay were evaluated in 0.5, 1.5, 3, 6, 24 hours after treatment. The cell proliferation and cell activity was also evaluated at 2 and 7 days after 1 minute exposure, twice a day. The cell morphology was similar between the Lipomyces smkeyi KSM 22 glucanhydrolase groups and control group during the incubation periods, while most fibroblasts remained as round cell regardless of incubation time in the chlorhexidine group. The numbers of the attached cells in the glucanhydrolase groups were comparable to that of control and significantly higher than the chlorhexidine group. The numbers of the proliferated cells in the glucanhydrolase groups at 7 days of incubation were comparable to the control group and higher than the chlorhexidine group. The cell activity in glucanhydrolase groups paralleled with the increased cell number by attachment and proliferation. According to these results, Lipomyces starkeyj KSM 22 glucanhydrolase has little harmful effect on attachment and proliferation of human gingival fibroblasts, in contrast to 0.1% chlorhexidine which was cytotoxic to human gingival fibroblasts. Therefore this glucanhydrolase preparation is considered as a safe and promising agent for new mouthwash formula in the near future.

• KSBB Journal
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• v.14 no.6
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• pp.713-717
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• 1999

We have isolated a dextranase and amylase constitutive and hyper-producing mutant, Lipomyces starkeyi JLC26, from Lipomyces starkeyi ATCC74054 after mutation using UV irradiation. After partial purification of dextranase and amylase (together DXAMase;both activities were always co-purified) by ammonium sulfate precipitation, CM-Sepharose column chromatography, the specific activities of amylase and dextranase were 5367 and 3045 unit/mg, respectively. The pH effects for activity and stabiligy of both enzymes were similar to each other: Optimum pH and temperature for activity sere at 5.5 and 37$^{\circ}C$ and optimum ranges for stability were at pH 2.5-5.5 and 4-55$^{\circ}C$, respectively. The reaction end products of dextranase and amylase activities were found to the typical for those of endo-dextranase and endo-amylase. When the carbohydrase and maltotriose were reacted, glucose, maltose, isomaltose, maltotriose, panose and ${\alpha}(1{\rightarrow}6)$glucosylmaltotriose were produced by disproportionation reaction.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.195-204
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• 2004

A novel glucanhydrolase from Lipomyces starkeyi KSM 22 has been suggested as a promising anti-plaque agent because it has been shown to have additional amylase activity and mutanase activity besides dextranase activity and to strongly bind to hydroxyapatite. Mouthrinsing with Lipomyces starkeyi KSM 22 glucanhydrolase solution was comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effects were less frequent and less intense in human experimental gingivitis. In this study, Lipomyces starkeyi KSM 22 glucanhydrolase mouthrinses (1 and 2 unit/ml) were compared with a control mouthrinse (commercial 0.01% benzethonium chloride mouthrinse, $Caregargle^{(R)}$, Hanmi Pharmaceuticals) in the ability to inhibit plaque formation. A 3-replicate clinical trial using 4-day plaque regrowth model was used. Fifteen volunteers were rendered plaque-free on the 1st day of each study period, ceased toothcleansing, and rinsed 2X daily with allocated mouthrinse thereafter. On day 5, plaque accumulation was scored and the washout periods was 9 days for the next trial. Lipomyces starkeyi KSM22 glucanhydrolase(1 unit and 2 unit)- containing mouthrinse resulted in Significantly lower plaque formation in plaque area and thickness, compared to the control mouthrinse. There was no significant difference in plaque inhibition between enzyme-mouthrinses at 2 different concentrations used. This glucanhydrolase- containing mouthwash resulted in significantly lower plaque area severity index score and tended to have lower plaque thickness severity index score than those of control mouthrinse. But there was no significant difference according to the enzyme concentration. From these results, Lipomyces starkeyi KSM 22 glucanhydrolase-containing benzethonium chloride mouthrinse has greater anti-plaque effect than the commercial mouthrinse alone. Therefore this glucanhydrolase preparation is a promising agent for new mouthwash formulation in the near future.

• Microbiology and Biotechnology Letters
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• v.27 no.4
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• pp.304-310
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• 1999

We have developed a new process for the production of new structure oligosaccharides using the mixed-culture fementation of Lipomyces starkeyi KSM22 and leuconostoc mesenteroides B-512FMCM.L.starkeyi KSM22 produces a novel DXAMase(an enzyme containing both dextranase and amylase activities). It hydrolyzes the soluble starch and dextran. The hydrolyzates were used as acceptors for dextransucrase of L.mesenteroides to synthesize the new oligosaccharides(NOS). In fermentation, as the concentration of sucrose was increased from 9%(w/v) to 15%(w/v), the yields of dextran(sum of dextran I, MW=66kD, and dextran II, MW=21kD) was increased from 12.7% to 42.5%, and NOS was increased from 3.9% to 5.2% of the theoretical, respectively. The NOS of dp(degree of polymerization) 5 and over was increased from 33.1% to 58.3% of the total NOS. The NOS showed heat resistant up to 12$0^{\circ}C$ and was stable at pHs ranged from 2 to 6. The NOS decreased the pH changes in the culture of S. mutans, and also showed inhibitory effects on the growth of S. aureus or S. typhimurium.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.700-703
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• 2001

The influence of process conditions on highly branched glucooligosaccharides production by mixed culture of Leuconostoc mesenteroides ATCC 13146 and Lipomyces starkeyi ATCC 74054 was studied. We divided the batch culture fermentations into two groups according to inoculation method. One-point inoculation was performed by coinoculation of L. mesenteroides and L. starkeyi at the ration of 10 to 1, and two-point inoculation by L. mesenteroides inoculation first and L. starkeyi inoculation after L. mesenteroides grew to the end of the log phase of growth. Two-point inoculation improved the yield of oligosaccharide by 1.5 to 20 fold more than one-point inoculation. In this process, the highest yield of oligosaccharides (48% of theoretical yield) and productivity (0.85 g/l/h) were obtained with starch as an initial substrate for L. starkeyi growth. The estimated composition of the end product consisted of 31.5% oligosaccharides, 17.6% dextran, and 46.5% mannitol.

• Journal of Life Science
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• v.19 no.4
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• pp.457-461
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• 2009

We optimized dextranase culture conditions by batch fermentation using Lipomyces starkeyi KCTC 17343. Furthermore, dextranase was purified by an ultra-membrane, and then dextran hydrolyzates were characterized. Cell growth and dextranase production varied depending on the initial culture pH and temperature. The conditions of optimal dextranase production were met in a pH range of 4-5 and temperature between $25-30^{\circ}C$. At optimal fermentation conditions, total enzyme activity and specific enzyme activity were about 4.85 IU/ml and 0.79 IU/g cells, respectively. The specific growth rate was examined to be $0.076\;hr^{-1}$. The production of dextranase in culture broth was very stably maintained after mid-log phase of growth. The enzyme hydrolyzed dextran into DP (degree of polymerization) 2 to 8 oligodextran series. Analysis of the composition of hydrolysates suggested that the enzyme produced is an endo-dextranase.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.983-987
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• 2006

The sole use of the glucanhydrolase (exhibiting both dextranase and amylase activities) from Lipomyces starkeyi hydrolyzed the soluble polysaccharides in sugar syrup more efficiently than a mixed treatment using both commercial dextranase and amylase. The glucanhydrolase treatment of stale sugar cane juice resulted in a yield of square, light-colored sugar crystals.

• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.993-997
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• 2002

A glucanhydrolase (a DXAMase exhibiting both dextranolytic and amylolytic activities) from Lipomyces starkeyi KSM 22 hydrolyzed polysaccharides having ${\alpha}-(1{\rightarrow}3)-,\;{\alpha}(1{\rightarrow}4)-,\;and\;{\alpha}-(1{\rightarrow}6)$-D-glucosidic linkages. The oral hygiene benefits of DXAMase-containing mouthwash were examined in relation to human experimental gingivitis during a 3-week period without brushing. The DXAMase-treated group exhibited a lower increase in plaque accumulation and gingival index score than the chlorhexidine-treated group. The DXAMase-treated group also showed less tongue accumulation, bad taste, and tooth staining, thus indicating a positive role for DXAMase as an antiplaque agent ingredient.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.313-316
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• 2003

Lipomyces starkeyi KSM 22 elaborates an enzyme that has both dextranase and amylase activities in a single protein of 100 kDa. Competition studies, using different amounts of dextran and starch as substrates, gave a competition plot consistent with the hypothesis that the hydrolysis of dextran and starch occurs at two independent active sites, each specific for starch and dextran, respectively.