• 한국식품과학회지
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• 제36권1호
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• pp.116-122
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• 2004

L. acidophilus를 면 제조 시에 첨가할 목적으로 MRS broth 배지를 이용하여 $37^{\circ}C$에서 3일 동안 배양시킨 후 면의 주원료인 밀가루, 물, 소금의 혼합액에 L. acidophilus를 선택 접종시켜 72시간 배양 발효시킨 결과 발효물에 젖산이 6.821mg/g, 초산이 0.191 mg/g 생성되어 발효물에서 젖산의 함량이 높았다. 배양기간 중 발효물의 pH는 저하되며 총산도가 증가되었다. 식염첨가 발효물의 점도는 발효시간의 경과에 따라 증가되었으나, 식염을 첨가하지 않은 발효물의 점도는 감소하였다. 한편 식염을 첨가하지 않은 발효물은 변질이 되어 식염이 보존성에 미치는 효과를 알 수 있었고 형태적 관찰에서 발효물에서 L. acidophilus의 전형적인 간균 형태를 보이고 있음을 확인할 수 있었다.

• 한국미생물·생명공학회지
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• 제19권6호
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• pp.598-603
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• 1991

신규 미생물 Bacillus subtilis subsp. krictiensis의 배양액으로부터 식물 및 인체 병원균에 폭넓은 항진균 활성을 나타내는 DRF-001으로 총칭되는 6개의 cyclic peptide의 복합체(A에서 F)를 분리하였다. 이 6개 peptides의 분자량은 FAB-MS 측정결과, A가 1042, B와 C가 1056, D와 E가 1070 그리고 F가 1084였다. 이 복합체의 구조를 각종 자기분석을 통해 해석한 결과, 구조 중에 1몰씩의 glutamine, proline, tyrosine, serine 및 unusal $\beta$-amino acid와 3몰의 asparagine을 공통적으로 갖고 있으나, $\beta$-amino acid의 탄소수와 말단 methyl 구조에 차이점이 있음을 알 수있었다. 구성 amino acid들의 서열과 입체구조 결정에 의하여 KRF-001의 잠정구조를 결정하였다.

• Applied Biological Chemistry
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• 제39권1호
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• pp.16-19
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• 1996

청주의 주질을 향상시키기 위한 전보의 연구에서, 유용효모로서 분리된 균주에 대하여 균학적 성질을 조사하고, 일부의 사항에 대하여는 일본청주효모와 그 특성을 비교하였다. 분리효모 KP-16, KP-21 및 KP-54 균주는 모두 saccharomyces cerevisae로 동정되었고, TTC 염색성이 pink계이었다. 맥아즙 배지에서의 피막(被膜) 형성능은 KP-16 및 KP-21 균주는 약하였으나, KP-54균주는 강하였다. 당의 발효성과 탄소원의 자화성은 KP-16과 KP-21 균주는 동일하였고, KP-54 균주는 다소 달랐다. 분리효모는 모두 ${\alpha}-methyl-D-glucoside$를 발효도 자화도 하지 못하였으며, 이는 현재 사용되고 있는 일본청주효모와 다른 점 중의 하나였다. 비타민 요구성에 있어서 분리균주는 모두 biotin과 pantothenate를 요구하였으며, biotin을 요구하는 점은 일본청주효모와는 다른 특성이었다. 내알코올성은 분리균주 모두 일본청주효모 K-7및 K-9균주보다 강하였다.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.187-192
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• 1988

Cholesterol로부터 미생물전환 방법에 의한 4-androstene-3,17-dione(AD)의 생성에 대해 연구를 수행했다. 발효액중에 cholesterol의 용해도를 증가시키기 위해 ethanol을 용매로 사용하였는데, ethanol 농도가 2%(v/v)까지는 세균성장이 크게 저해되지 않았다. 미생물전환은 pH를 7.0으로 조절하고, 초기대수 증식기에 cholesterol을 첨가했을 때 효율적으로 AD가 생성되었다. AD 생성을 높이기 위해 cholesterol 첨가방법을 여러 가지로 변환시켰다. 즉, 최종 cholesterol 농도를 0.1% 하여 ethanol에 녹여 간헐적으로 첨가했을 때 가장 높은 수율을 얻었다. Cholesterol을 세번(전체 3g/$\ell$) 첨가했을 때 최종 전환수율이 65%에 도달한 반면, 같은 양의 cholesterol (3g/$\ell$)을 한번에 넣었을 때 40%의 생성수율을 얻었다.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1742-1750
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• 2015

Human insulin is composed of 21 amino acids of an A-chain and 30 amino acids of a B-chain. This is the protein hormone that has the role of blood sugar control. When the recombinant human proinsulin is expressed in Escherichia coli, a serious problem is the formation of an inclusion body. Therefore, the inclusion body must be denatured and refolded under chaotropic agents and suitable reductants. In this study, H27R-proinsulin was refolded from the denatured form with β-mercaptoethanol and urea. The refolding reaction was completed after 15 h at $15^{\circ}C$, whereas the reaction at $25^{\circ}C$ was faster than that at $15^{\circ}C$. The refolding yield at $15^{\circ}C$ was 17% higher than that at $25^{\circ}C$. The refolding reaction could be carried out at a high protein concentration (2 g/l) using direct refolding without sulfonation. The most economical and optimal refolding condition for human preproinsulin was 1.5 g/l protein, 10 mM glycine buffer containing 0.6 M urea, pH 10.6, and 0.3 mM β-mercaptoethanol at $15^{\circ}C$ for 16 h. The maximum refolding yield was 74.8% at $15^{\circ}C$ with 1.5 g/l protein. Moreover, the refolded preproinsulin could be converted into normal mature insulin with two enzymes. The average amount of human insulin was 138.2 g from 200 L of fermentation broth after enzyme reaction with H27R-proinsulin. The direct refolding process for H27R-proinsulin was successfully set up without sulfonation. The step yields for refolding and enzyme reaction were comparatively high. Therefore, our refolding process for production of recombinant insulin may be beneficial to the large-scale production of other biologically active proteins.

• 한국미생물·생명공학회지
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• 제40권4호
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• pp.303-309
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• 2012

Keratinases are of particular interest because of their action on insoluble keratins and generally on a broad range of protein substrates. Alkalophilic and neutrophilic actinomycete strains isolated from different soil samples, rich in keratinaceous substances were screened for keratinolytic activity. An alkalophilic isolate, TBG-S13A5, was found to possess good keratinolytic activity and was able to utilize feather as the sole carbon and nitrogen source. TBG-S13A5 exhibited an off-white aerial mass color with a rectus-flexibilis type of spore chain. The morphological, microscopical and biochemical characters were comparable with that of Streptomyces albidoflavus. Fatty acid methyl ester profiling (FAME) and 16S rDNA sequence analysis confirmed its identity as a strain of S. albidoflavus. Under submerged fermentation conditions, maximum protease production was recorded on the $5^{th}$ day of incubation at $30^{\circ}C$, using basal broth of pH 9.0 with 0.25% (w/v) white chicken feather. This strain could affect feather degradation when the initial pH was 8 and above and maximum protease production was recorded when the initial pH was around 10.5. The effectiveness of the crude enzyme in destaining and leather dehairing were also demonstrated.

• The Korean Journal of Physiology and Pharmacology
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• 제16권2호
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• pp.145-151
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• 2012

In the present work, we studied the structure-activity relationship (SAR) of tautomycetin (TMC) and its derivatives. Further, we demonstrated the correlation between the immunosuppressive fuction, anticancer activity and protein phosphatase type 1 (PP1) inhibition of TMC and its derivatives. We have prepared some TMC derivatives via combinatorial biosynthesis, isolation from fermentation broth or chemical degradation of TMC. We found that the immunosuppressive activity was correlated with anticancer activity for TMC and its analog compounds, indicating that TMC may home at the same targets for its immunosuppressive and anticancer activities. Interestingly, TMC-F1, TMC-D1 and TMC-D2 all retained significant, albeit reduced PP1 inhibitory activity compared to TMC. However, only TMC-D2 showed immunosuppressive and anticancer activities in studies carried out in cell lines. Moreover, TMC-Chain did not show any significant inhibitory activity towards PP1 but showed strong growth inhibitory effect. This observation implicates that the maleic anhydride moiety of TMC is critical for its phosphatase inhibitory activity whereas the C1-C18 moiety of TMC is essential for the inhibition of tumor cell proliferation. Furthermore, we measured $in$ $vivo$ phosphatase activities of PP1 in MCF-7 cell extracts treated with TMC and its related compounds, and the results indicate that the cytotoxicity of TMC doesn't correlate with its $in$ $vivo$ PP1 inhibition activity. Taken together, our study suggests that the immunosuppressive and anticancer activities of TMC are not due to the inhibition of PP1. Our results provide a novel insight for the elucidation of the underlying molecular mechanisms of TMC's important biological functions.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.71-76
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• 2002

Soil samples were screened for actinomycete strains capable of producing phospholipase D, and a strain, Streptomyces sp. YU100, showing a high transphosphatidylation activity was isolated. This strain secreted phospholipase D in a culture broth after 12 h of cultivation, and its productivity continued to increase for 36 h of fermentation. In addition, its transphosphatidylation rate of phosphatidylcholine to phosphatidylserine was almost $68\%$ within 1 h. The morphological and chemotaxonomical characteristics showed that this strain could be classified as a number of the Streptomycetaceae family, particularly due to the spiral form of its spore chain consisting of 60-70 smooth spores $(0.75{\times}1.0{\mu}m$) on an aerial mycelium, FA-2c type of fatty acid profile in the cell wall, and LL-DAP component in the cell wall peptidoglycan. A phylogenetic analysis of the 16S rDNA provided a clue that the strain YU100 was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyes sp. ASB27, S. peucetius JCM9920, and S. griseus ATCC10137. A dendrogram based on the 16S rDNA sequences also showed a phylogenetic relationship between the strain YU100 and these strains. However, the strain YU100 has not yet been assigned to a particular species, because of absence of any other classified species with a high matching score.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1379-1385
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• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1500-1512
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• 2017

In this study, the siderophore-producing characteristics and conditions of Bacillus sp. PZ-1 were investigated and the enhancement of siderophores on Pb uptake and translocation in Brassica juncea were determined. Results of single factor experiment showed that glucose, pH, and $Pb(NO_3)_2$ could stimulate PZ-1 growth and siderophore production. The maximum siderophore production of 90.52% siderophore units was obtained by response surface methodology optimization at the glucose concentration of 21.84 g/l, pH 6.18, and $Pb(NO_3)_2$ concentration of $245.04{\mu}mol/l$. The type of siderophore was hydroxamate and its concentration in the fermentation broth amounted to $32.24{\mu}g/ml$. Results of pot experiments indicated that the siderophores enhanced B. juncea to assimilate more Pb from soil with the uptake ratio from 1.04 to 2.74, and to translocate more Pb from underground to overground with the TF values from 1.21 to 1.48. The results revealed that Bacillus sp. PZ-1 could produce abundant siderophores and might be potentially used to augment the phytoextraction of Pb from soil.