• 동아시아식생활학회지
• /
• 제22권4호
• /
• pp.473-479
• /
• 2012

본 연구에서는 헛개나무 열매 추출물을 당 분해효소를 이용하여 효소처리 시 알코올 분해능에 미치는 영향을 알아보았다. 당분해효소인 Maxinvert(invertase), Optidex L-400(glucoamylase), Rohament CL(cellulase & pectinase)를 각각 농도별(0.01, 0.05, 0.1, 0,5, 1%)로 첨가하여 48시간 효소처리 하면서 6시간 간격으로 sample을 채취해 ADH 효소 활성을 측정하였다. 효소 농도가 높을수록 효소 활성이 증가하였으며, 시간이 지남에 따라 활성이 높아졌다. Maxinvert 효소 처리 시 기존의 활성보다 약 10% 증가하여 가장 낮은 활성 증가를 보였고, Rohament CL 효소를 1% 첨가하여 효소처리할 경우, 기존의 활성에 비해 효소 활성이 67% 증가한 것으로 가장 많은 효소 활성 증가를 보였다. Rohament CL 효소를 1% 첨가할 경우, 시간이 지남에 따라 활성이 증가하다가 30시간대부터 활성의 증가가 변화가 거의 없었다. 이로 보아 36시간까지 효소처리하는 것이 가장 효율적일 것이라 사료된다. Rohament CL 효소 1% 첨가하여 36시간 효소 처리한 헛개나무 열매 추출물을 이용하여 동물 실험 및 임상 실험을 실시한 결과, control군 및 헛개나무 열매 추출물 군에 비해 알코올 농도가 줄어드는 것을 볼 수 있었으며, 특히 알코올 섭취 초기에 그 효과가 가장 뛰어난 것으로 나타났다. 이로 보아 효소 처리한 헛개나무 열매 추출물이 기존의 헛개나무 열매 추출물에 비해 더 많은 체내 알코올 농도 감소를 나타낼 것으로 생각되며, 음주 초기에 더 많은 영향을 줄 것으로 사료되어 진다. 이는 기존의 배당체(glycoside) 형태로 존재하던 생리활성 물질이 당 분해 효소에 의해 당이 분해된 무배당체(aglycone) 형태로 전환되어 그 활성이 높아졌기 때문으로 사료되어진다.

• Journal of Microbiology and Biotechnology
• /
• 제24권11호
• /
• pp.1525-1535
• /
• 2014

The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

• Mycobiology
• /
• 제48권2호
• /
• pp.104-114
• /
• 2020

The carbohydrate-active enzyme (CAZyme) genes of Trametes contribute to polysaccharide degradation. However, the comprehensive analysis of the composition of CAZymes and the biosynthetic gene clusters (BGCs) of Trametes remain unclear. Here, we conducted comparative analysis, detected the CAZyme genes, and predicted the BGCs for nine Trametes strains. Among the 82,053 homologous clusters obtained for Trametes, we identified 8518 core genes, 60,441 accessory genes, and 13,094 specific genes. A large proportion of CAZyme genes were cataloged into glycoside hydrolases, glycosyltransferases, and carbohydrate esterases. The predicted BGCs of Trametes were divided into six strategies, and the nine Trametes strains harbored 47.78 BGCs on average. Our study revealed that Trametes exhibits an open pan-genome structure. These findings provide insights into the genetic diversity and explored the synthetic biology of secondary metabolite production for Trametes.

• Journal of Microbiology and Biotechnology
• /
• 제20권10호
• /
• pp.1403-1414
• /
• 2010

Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as an extracellular enzyme under submerged culture conditions. Enzymes with a specific activity of 2,761.89 IU/mg protein, a final yield of 0.51%, and a purification fold of 6.32 were obtained after purification through to homogeneity, by ultrafiltration and gel filtration. SDS-PAGE analyses, under nonreducing and reducing conditions, yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating the presence of six identical monomers. A pI of 4.4 and a carbohydrate content of 8.02% were observed in the enzyme. The optimal temperature was found to be $30^{\circ}C$, although the enzyme was active in the range of $5-80^{\circ}C$. Two pH optima, pH 2 and pH 8, were recorded, although the enzyme was instable at a pH of 8, but stable at a pH of 2.0 for 24 h. Methylgallate recorded maximal affinity, and $K_m$ and $V_{max}$ were recorded at $1.9{\times}10^{-3}$M and 830 ${\mu}Mol$/min, respectively. The impacts of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on tannase activity were determined in order to establish the novel characteristics of the enzyme. The gene encoding tannase, isolated from A. awamori, was found to be 1.232 kb, and nucleic acid sequence analysis revealed an open reading frame consisting of 1,122 bp (374 amino acids) of one stretch in the -1 strand. In silico analyses of gene sequences, and a comparison with reported sequences of other species of Aspergillus, indicate that the acidophilic tannase from marine A. awamori differs from that of other reported species.

• Applied Biological Chemistry
• /
• 제13권1호
• /
• pp.29-34
• /
• 1970

우수한 영양가를 가진 대두를 이용하여 이유식을 제조하기 위하여 가압증자한 대두에 Protease및 Cellulase 역가가 비교적 높은 Asp. niger 및 Asp. sojae균의 피국 추출조효소액을 작용시켜 대두 단백질을 아미노산 내지 Peptide 태로 분해시키는 최적조건을 결정하고 여기서 얻은 분해물을 탈색 농축시키는 효과에 관하여 실험하여 다음과 같은 결과를 얻었다. 1. 대두의 가압증자는 15Ib에서 10분간 처리함이 가장 높은 단백응해율 및 단백분해율을 나타냈다. 2. Asp. sojae enzyme은 pH 6.0, Asp. niger enzyme은 pH 4.4에서 가장 높은 단백용해율과 단백분해율을 나타냈다. Asp. sojae enzyme을 처리한 다음 Asp. niger emzyme을 작용 분해시킨 것이 각 효소 단독으로처리했을때 보다 높은 단백응해율(62.3%) 및 단백분해율을 (56.4%) 나타냈다. 3. 기질에 대한 효소액 첨가량은 원료 대두에 10배의 물을 가한 마쇄기질액에 피국에 대하여 10배의 물로 추출한 효소액 1/2에 해당하는 양 (Asp. sojae enzyme와 Asp. niger emzyme 총량)을 가하는것이 가장 실용적이었다. 4. 분해시간이 길 수록 단백응해율 및 단백분해율이 높아지나 부패 등을 고려할 때 실용분해 시간은 8시간 정도가 적당하다. 5. 탈색 효과는 활성탄으로 처리한 후 음이온교환수지 (Dowex 2-x-8)을 처리한 것이 가장 좋고 단독처리로는 음이온교환수지, 활성탄, 양이온교환 수지(Amberite)의 순으로 효과가 적었다. 6. 이상의 최적 조건으로 대두단백질을 분해하고 $60^{\circ}C$ 이하에서 감압농축하여 얻은 제품은 수분 12.51%, 단백질 66.31%, 지방 4.25%, 탄수화물 12.75%,인 건조분말을 얻었다. 이 연구는 1969년도 문교부 학술연구 조성비로 이루워진 것이며 본 연구에 헌신적인 보조를 아끼지 않았던 박 관화 군에게 감사하는 바이다.

• Journal of Microbiology and Biotechnology
• /
• 제29권8호
• /
• pp.1255-1265
• /
• 2019

To investigate the diversity of gastrointestinal microflora and lignocellulose-degrading enzymes in wild Asian elephants, three of these animals living in the same group were selected for study from the Wild Elephant Valley in the Xishuangbanna Nature Reserve of Yunnan Province, China. Fresh fecal samples from the three wild Asian elephants were analyzed by metagenomic sequencing to study the diversity of their gastrointestinal microbes and cellulolytic enzymes. There were a high abundance of Firmicutes and a higher abundance of hemicellulose-degrading hydrolases than cellulose-degrading hydrolases in the wild Asian elephants. Furthermore, there were a high abundance and a rich diversity of carbohydrate active enzymes (CAZymes) obtained from the gene set annotation of the three samples, with the majority of them showing low identity with the CAZy database entry. About half of the CAZymes had no species source at the phylum or genus level. These indicated that the wild Asian elephants might possess greater ability to digest hemicellulose than cellulose to provide energy, and moreover, the gastrointestinal tracts of these pachyderms might be a potential source of novel efficient lignocellulose-degrading enzymes. Therefore, the exploitation and utilization of these enzyme resources could help us to alleviate the current energy crisis and ensure food security.

• Journal of Microbiology and Biotechnology
• /
• 제27권3호
• /
• pp.591-597
• /
• 2017

Maribacter dokdonensis DSW-8 was isolated from the seawater off Dokdo in Korea. To investigate the genomic features of this marine bacterium, we sequenced its genome and analyzed the genomic features. After de novo assembly and gene prediction, 16 contigs totaling 4,434,543 bp (35.95% G+C content) in size were generated and 3,835 protein-coding sequences, 36 transfer RNAs, and 6 ribosomal RNAs were detected. In the genome of DSW-8, genes encoding the proteins associated with gliding motility, molybdenum cofactor biosynthesis, and utilization of several kinds of carbohydrates were identified. To analyze the genomic relationships among Maribacter species, we compared publically available Maribacter genomes, including that of M. dokdonensis DSW-8. A phylogenomic tree based on 1,772 genes conserved among the eight Maribacter strains showed that Maribacter speices isolated from seawater are distinguishable from species originating from algal blooms. Comparison of the gene contents using COG and subsystem databases demonstrated that the relative abundance of genes involved in carbohydrate metabolism are higher in seawater-originating strains than those of algal blooms. These results indicate that the genomic information of Maribacter species reflects the characteristics of their habitats and provides useful information for carbon utilization of marine flavobacteria.

• Journal of Microbiology and Biotechnology
• /
• 제28권12호
• /
• pp.2036-2045
• /
• 2018

An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

• 한국식품영양학회지
• /
• 제33권3호
• /
• pp.309-316
• /
• 2020

In this study, we investigated the protective effects of Ulva lactuca methanol extracts against ultraviolet B (UVB)-induced DNA damage in HaCaT cells. First, the contents of general and antioxidative nutrient contents of Ulva lactuca were measured. The moisture, carbohydrate, crude protein, crude fat and ash were 14.01%, 44.80%, 23.19%, 3.10% and 14.90%, respectively. Magnesium that acts as DNA repair enzyme cofactor was the most abundant mineral followed by Ca, P and Fe. The total phenolic and anthocyanoside contents of Ulva lactuca were 2.69 mg/g and 0.13 mg/g, respectively. Cells treated with Ulva lactuca methanol extracts for 24 hours post UVB exposure increased cell viability in a concentration-dependent manner compared to the non-treated control. Also, Ulva lactuca methanol extracts decreased the levels of UVB-induced DNA damage such as cyclobutane pyrimidine dimer and DNA damage response (DDR) proteins such as p-p53 and p21. These results suggest that Ulva lactuca methanol extracts comprising physiological active substances such as Mg, polyphenols and anthocyanosides promote DNA repair by regulating genes related with DDR.

• 미생물학회지
• /
• 제54권4호
• /
• pp.463-464
• /
• 2018

Tamlana sp. UJ94는 해수로부터 분리되었으며 알긴산을 분해할 수 있다. 알긴산 분해 관련 특성을 이해하기 위해 이 세균의 유전체를 분석하였다. UJ94의 유전체는 4,116,543 bp의크기로 3,609개의 코딩서열을 가지고 있으며 35.2 mol%의 G + C 함량을 가진다. BLASTp 검색 결과 9개의 alginate lyase 외에도 6개의 agarase, 5개의 amylase, 4개의 carrageenase, 1개의 cellulase, 4개의 pectate lyase, 7개의 xylanase의 존재가 예측되어 UJ94의 다양한 다당류 분해 능력을 암시하였다. Tamlana sp. UJ94의 유전체는 생물전환 공정에 사용할 수 있는 다당류 분해 유전자를 제공할 수 있을 것이다.