Indonesian Journal of Biotechnology最新文献_第9页

Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids CRISPR/Cas9基因组编辑系统在兰花分子育种中的应用

Indonesian Journal of Biotechnology Pub Date : 2020-06-30 DOI: 10.22146/ijbiotech.39485

E. Semiarti, S. Nopitasari, Y. Setiawati, M. D. Lawrie, A. Purwantoro, J. Widada, Y. Yoshioka, S. Matsumoto, K. Ninomiya, Yuuki Asano

{"title":"Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids","authors":"E. Semiarti, S. Nopitasari, Y. Setiawati, M. D. Lawrie, A. Purwantoro, J. Widada, Y. Yoshioka, S. Matsumoto, K. Ninomiya, Yuuki Asano","doi":"10.22146/ijbiotech.39485","DOIUrl":"https://doi.org/10.22146/ijbiotech.39485","url":null,"abstract":"Orchid is an important ornamental plant in Indonesia due to their natural beauty of flowers. In the tropical forest, orchids are being acquired for trading and commercial market. Thus, the effort is required to proliferate orchid in large quantities for conservation and improve the floral variation for plant breeding. The purpose of this study is to develop a firmed methodology of molecular breeding of orchids using CRISPR/Cas9 KO system. The plant material used was Phalaenopsis amabilis protocorms growth on NP medium+pepton (2 g/L). Protocorm were submerged in the culture of Agrobacterium tumefaciens that Ti‐plasmid had been filled with a T‐DNA construct of a pRGEB32 vector harboring sgRNA with PDS3 sequence. Detection for transformants was confirmed by PCR using HPT primers (545 bp), Cas9 primers (402 bp), PDS primers (280 bp) and trnL‐F (1200 bp) as an internal control. The results showed that 0.96% PDS transformants were obtained from PDS3T2 lines. Several transformant showed pale leaf color compared to non‐transformant plants. This study suggests that the target gene has successfully edited by CRISPR/Cas9 system and could be applied for that functional gene editing in orchids.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"61-68"},"PeriodicalIF":0.0,"publicationDate":"2020-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49192701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Chitosan suppresses the expression level of WRKY17 on red chili (Capsicum annuum) plant under drought stress 壳聚糖抑制干旱胁迫下红辣椒WRKY17基因的表达水平

Indonesian Journal of Biotechnology Pub Date : 2020-06-29 DOI: 10.22146/ijbiotech.55016

M. Aziz, R. Esyanti, K. Meitha, F. Dwivany, Hany Husnul Chotimah

{"title":"Chitosan suppresses the expression level of WRKY17 on red chili (Capsicum annuum) plant under drought stress","authors":"M. Aziz, R. Esyanti, K. Meitha, F. Dwivany, Hany Husnul Chotimah","doi":"10.22146/ijbiotech.55016","DOIUrl":"https://doi.org/10.22146/ijbiotech.55016","url":null,"abstract":"Chili pepper plays a significant role in the global market. However, the production is often impeded by drought stress involving WRKY genes as the defense regulator. Chitosan is considered as a promising alternative fertilizer and defense elicitor. Hence, this study aimed to determine the role of chitosan in improving plant growth and survival of red chili pepper against drought stress. At the onset of the generative phase, chili plants were subjected to 1 mg mL‐1 chitosan, 50 percent drought, or chitosan‐drought treatment. Observations were made on several growth parameters, opened stomata, and WRKY gene expression. The results showed that chitosan‐drought treatment decreased plant growth and yielded significantly. The percentage of opened stomata was recorded at 0.56‐fold lower than control. It was followed by the decrease of the relative expression of WRKY17 and WRKY53 genes up to 0.56 and 0.72‐fold lower than control, respectively. Therefore, we suggested that the double treatment of chitosan‐drought might decrease plant growth performance but increase the defense system by suppressing the expression level of the WRKY17 gene. Interestingly, the drought treatment significantly increased WRKY17 expression level up to 7‐fold higher than control. Hence, it was suggested that WRKY17 has a specific role in response to drought stress.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"52-60"},"PeriodicalIF":0.0,"publicationDate":"2020-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42996952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Differentiation ability of rat‐mesenchymal stem cells from bone marrow and adipose tissue to neurons and glial cells 大鼠骨髓和脂肪组织间充质干细胞向神经元和神经胶质细胞的分化能力

Indonesian Journal of Biotechnology Pub Date : 2020-06-26 DOI: 10.22146/ijbiotech.42511

Ariyani Noviantari, Ratih Rinendyaputri, Ibnu A. Ariyanto

{"title":"Differentiation ability of rat‐mesenchymal stem cells from bone marrow and adipose tissue to neurons and glial cells","authors":"Ariyani Noviantari, Ratih Rinendyaputri, Ibnu A. Ariyanto","doi":"10.22146/ijbiotech.42511","DOIUrl":"https://doi.org/10.22146/ijbiotech.42511","url":null,"abstract":"Mesenchymal stem cells (MSCs) are multipotent cells and can differentiate into neurons and glial cells. In vitro differentiation would be done by the addition of various factors. There remains no comparison for the differentiation of MSCs from rat bone marrow (rBMMSCs) and adipose tissue (rATMSCS) into neurons and glial cells with basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and brain‐derived neurotrophic factor (BDNF). The aims of this study were to investigate the effect of bFGF, EGF, and BDNF supplementation on the differentiation ability of rBMMSCs and rATMSCs into neurons and glial cells. MSCs were cultured with bFGF and EGF for 4 days and then BDNF was added until day 8. Characterization of MSCs before and after induction was carried out by observing the cell morphology and several cell markers. Flowcytometry analysis was performed for MSCs markers (CD90, CD29) and neurons and glial cell markers (A2B5, Beta‐III‐tubulin, PSAN‐CAM); while MAP‐2, a neuron marker, was analyzed by immunocytochemistry. Induction of both types of MSCs showed MAP‐2‐positive cells, decreased MSCs markers, and in rBMMSCs showed increased neuron markers. The number of neuron marker positive cells in rBMMSCS was higher than rATMSCs. This study showed that the addition of bFGF, EGF, and BDNF to the medium induced rBMMSCs into neurons and glial cells, but the conditions were not optimal for rATMSC as judged by the expression of neural markers (A2B5, Beta‐III‐tubulin, PSAN‐CAM, and MAP‐2).","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"43-51"},"PeriodicalIF":0.0,"publicationDate":"2020-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48787809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Enhancement of transient erythropoietin protein expression by valproic acid in CHO‐K1 suspension adapted cells 丙戊酸增强CHO‐K1悬浮液适应细胞中瞬时促红细胞生成素蛋白的表达

Indonesian Journal of Biotechnology Pub Date : 2020-06-18 DOI: 10.22146/ijbiotech.52621

Y. Rubiyana, R. Soejoedono, A. Santoso

{"title":"Enhancement of transient erythropoietin protein expression by valproic acid in CHO‐K1 suspension adapted cells","authors":"Y. Rubiyana, R. Soejoedono, A. Santoso","doi":"10.22146/ijbiotech.52621","DOIUrl":"https://doi.org/10.22146/ijbiotech.52621","url":null,"abstract":"Erythropoietin (EPO) is a therapeutic protein that is widely used to increase red blood cell production in chronic kidney failure. EPO protein can be produced quickly with a transient gene expression system (TGE). However, the titer produced using TGE is usually lower than the stable gene expression system (SGE). It has been known that TGE can be improved by histone deacetylase inhibitors (iHDACs) such as valproic acid (VPA). This study was conducted to examine the VPA effect on EPO protein expression in CHO‐K1 suspension adapted cells and to find the optimum concentration of VPA on transient EPO protein production. EPO proteins was quantified using the enzyme‐linked immunosorbent assay (ELISA) method. The optimization of VPA concentrations showed that VPA increased the EPO protein yield by up to 2‐fold in transient EPO production, and the optimum concentration of VPA was 4 mM. VPA optimization was very helpful to obtain the maximum increase in the transiently expressed protein. Furthermore, this study can be used as a model to produce EPO proteins or other recombinant proteins rapidly with TGE of CHO‐K1 suspension adapted cells.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47719208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The structural insight of class III of polyhydroxyalkanoate synthase from Bacillus sp. PSA10 as revealed by in silico analysis 通过硅分析揭示了芽孢杆菌PSA10聚羟基烷酸酯合酶III类的结构特征

Indonesian Journal of Biotechnology Pub Date : 2020-06-18 DOI: 10.22146/ijbiotech.53717

Listia Pradani, M. S. Rohman, S. Margino

{"title":"The structural insight of class III of polyhydroxyalkanoate synthase from Bacillus sp. PSA10 as revealed by in silico analysis","authors":"Listia Pradani, M. S. Rohman, S. Margino","doi":"10.22146/ijbiotech.53717","DOIUrl":"https://doi.org/10.22146/ijbiotech.53717","url":null,"abstract":"PhaC synthase is an enzyme responsible for PHA polymerization. In this work, the catalytic mechanism class III of PhaC synthase from Bacillus sp. PSA10 (BacPhaCSynt) was reported through in silico modelling approach based on the primary sequence of the PhaC synthase. The open reading frame BacPhaCSynt has been successfully isolated, cloned and overexpressed the recombinant protein in Escherichia coli BL21(DE3). To know the global architecture and catalytic mechanism, the structural prediction of BacPhaCSynt has been carried out by using MODELLER. The recombinant BacPhaCSynt exhibited monomeric molecular weight (MW) of 43.6 kDa, when it was analyzed on 12% SDS‐PAGE gel. Based on the structural prediction, BacPhaCSynt exhibited global architecture of α/β hydrolase fold, with the root mean square deviation (r.m.s.d) value of 0.94Å. The catalytic residues composition of BacPhaCSynt consists of C151, D307, and H336, but the H336 and D307 residues of the model have been distorted 62.8o and 175.2o from the corresponding residues of the template. Since the D307 is quite a distance from the H336, it might act as a general base for the activation of ‐OH group of the substrate. The results strongly suggested that the mode of action of BacPhaCSynt obeyed the covalent catalysis mechanism.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44370450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

The potency of Pentagamavunone‐0 (PGV‐0) as chemopreventive agent for the formation and growth of breast cancer as revealed in 3D model Pentagamavunone‐0 (PGV‐0)作为化学预防剂对乳腺癌形成和生长的效力在3D模型中揭示

Indonesian Journal of Biotechnology Pub Date : 2020-06-07 DOI: 10.22146/IJBIOTECH.51759

W. Wulandari, Muthi’ Ikawati, E. Meiyanto

{"title":"The potency of Pentagamavunone‐0 (PGV‐0) as chemopreventive agent for the formation and growth of breast cancer as revealed in 3D model","authors":"W. Wulandari, Muthi’ Ikawati, E. Meiyanto","doi":"10.22146/IJBIOTECH.51759","DOIUrl":"https://doi.org/10.22146/IJBIOTECH.51759","url":null,"abstract":"Pentagamavunone‐0 (PGV‐0) or 2,5‐bis(4’‐hydroxy‐3‐methoxybenzylidine)‐cyclopentanone is a curcumin analogue that exhibits anticancer activity in breast cancer cells. However, most of previous reports are limited to the use of two‐dimensional (2D) cell culture. The use of three‐dimensional (3D) cell culture model in cancer research can represent the real condition of cancer growth in patients better than the 2D culture. The purpose of this study was to determine the anticancer activity of PGV‐0 on a 3D model of HCC 1954 breast cancer cells. HCC 1954 cells were grown in the 3D culture in the presence of PGV‐0, and the spheroid formation and growth of formed spheroids were observed using microscope at 24 and 96 h, respectively. The cytotoxic effects were measured by MTT assay. PGV‐0 inhibited the formation and growth of spheroids at the concentration as low as 60 µM. The cytotoxic effect of PGV‐0 appeared in a dose‐dependent manner with the IC50 value of 70.9 µM. The results of this study indicate that PGV‐0 has an anticancer activity on a 3D model of HCC 1954 breast cancer cell line. Therefore, the result supported the potency of PGV‐0 as cancer chemopreventive agent.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"21-27"},"PeriodicalIF":0.0,"publicationDate":"2020-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46744377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Physiological, biochemical and HSP70 and HSP90 gene expression profiles of tropical abalone Haliotis squamata in response to Vibrio alginolyticus infection 热带鲍对溶藻弧菌感染反应的生理生化及HSP70和HSP90基因表达谱

Indonesian Journal of Biotechnology Pub Date : 2020-06-07 DOI: 10.22146/IJBIOTECH.51322

N. S. Yasa, M. Murwantoko, N. Handayani, G. Triastutik, L. Anshory

{"title":"Physiological, biochemical and HSP70 and HSP90 gene expression profiles of tropical abalone Haliotis squamata in response to Vibrio alginolyticus infection","authors":"N. S. Yasa, M. Murwantoko, N. Handayani, G. Triastutik, L. Anshory","doi":"10.22146/IJBIOTECH.51322","DOIUrl":"https://doi.org/10.22146/IJBIOTECH.51322","url":null,"abstract":"Vibrio spp. have been known responsible for fish diseases in marine and brackish‐water systems in the tropics regions. Heat shock proteins are a highly conserved protein group that is known for its rapid response to environmental stresses, including infection. This study aimed to investigate physiological and biochemical responses of tropical abalone Haliotis squamata to Vibrio alginolyticus infection. Abalones were infected with V. alginolyticus by intramuscular injection at a dose of 105,106,107 cfu/abalone. The expression of HSP70 and HSP90 genes, the activity of superoxide dismutase, phenol oxidase and catalase enzymes, histology, falling and mortality were observed at 12, 24, 48, 72, and 96 hours post‐infection (hpi). The different expression of HSPs was found in this study. While the expression of HSP70 was downregulated after infection, the expression of HSP90 was upregulated at 12 hpi and followed by downregulated after 24 hpi for 106 cfu infection, but expressed at a normal level for 105 infection treatment. The expression ofsuperoxide dismutase and catalase increased within 12 hpi, and the expression of phenol oxidase increased after 24 hpi. V. alginolyticus is virulent with LD50 of less than 105 cfu on H. squamata with an average weight of 5.13 g, and caused enlargement of hemolymph sinus and development intraepithelial and intramuscular abscesses.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"12-20"},"PeriodicalIF":0.0,"publicationDate":"2020-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47027338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Phylogenetic analysis of 23 accessions of Indonesian banana cultivars based on Internal Transcribed Spacer 2 (ITS2) region 基于ITS2区域的23份印尼香蕉品种系统发育分析

Indonesian Journal of Biotechnology Pub Date : 2020-05-18 DOI: 10.22146/IJBIOTECH.49506

K. Meitha, Intan Fatmawati, F. Dwivany, A. Sutanto, S. N. Pratama, H. Nugrahapraja, K. Wikantika

{"title":"Phylogenetic analysis of 23 accessions of Indonesian banana cultivars based on Internal Transcribed Spacer 2 (ITS2) region","authors":"K. Meitha, Intan Fatmawati, F. Dwivany, A. Sutanto, S. N. Pratama, H. Nugrahapraja, K. Wikantika","doi":"10.22146/IJBIOTECH.49506","DOIUrl":"https://doi.org/10.22146/IJBIOTECH.49506","url":null,"abstract":"Pisang Kepok ( Musa spp. [ABB ’Saba’ subgroup]) has several unique characteristics, such as tolerance to drought and Fusarium Foc (TR4) disease. Currently, the genetic diversity of Pisang Kepok in Indonesia is not well identified, although it is widely cultivated. Information on genetic diversity is essential for developing breeding strategies to achieve efficient cultivar improvement in the future. Aims of this research were to analyze the genetic variation of Pisang Kepok from some islands in Indonesia and to determine the genetic relationship between Pisang Kepok and other accessions banana cultivars based on ITS2 region, as a basis for future research in improving banana quality through molecular breeding. We have conducted the multiple sequence alignment and built the phylogenetic tree analysis using the Bayesian Inference Phylogeny method of one million generations (ngen = 1,000,000). The ITS2 region showed two clade ingroups: first clade consists of banana with B genome ( balbisiana ), while the second clade consists of banana with only A genome ( acuminata ). In general, all accessions of Pisang Kepok cultivars were clustered in the B genome of bananas cultivars. In addition, the ITS2 sequences and secondary structures among Pisang Kepok from various regions are identical, suggesting that there was no genetic variation in the ITS2 region of Pisang Kepok from multiple areas in Indonesia.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"25 1","pages":"1-11"},"PeriodicalIF":0.0,"publicationDate":"2020-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45034456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Expression profiling of the CHS8, CHI1A, IFS2, and CHR genes in black soybean seed [Glycine max (L). Merr.] of F4 generation 黑豆种子CHS8、CHI1A、IFS2和CHR基因的表达谱分析[j]。F4代的]

Indonesian Journal of Biotechnology Pub Date : 2020-01-03 DOI: 10.22146/ijbiotech.51888

D. Sumardi, Aulia Marwah Mumtaza, R. Maulani, A. Pancoro, H. Nugrahapraja, S. Suhandono, T. S. Syamsudin, A. Kurniawan

{"title":"Expression profiling of the CHS8, CHI1A, IFS2, and CHR genes in black soybean seed [Glycine max (L). Merr.] of F4 generation","authors":"D. Sumardi, Aulia Marwah Mumtaza, R. Maulani, A. Pancoro, H. Nugrahapraja, S. Suhandono, T. S. Syamsudin, A. Kurniawan","doi":"10.22146/ijbiotech.51888","DOIUrl":"https://doi.org/10.22146/ijbiotech.51888","url":null,"abstract":"Black soybean [ Glycine max (L.) Merr.] produces isoflavones as secondary metabolites, which have many benefits for human health and plant defense system. Expression profiling can guide potential work in functional genomics of the isoflavone biosynthesis pathway. Previous studies showed the vital role of the CHS8, CHI1A, and IFS 2 genes in isoflavone biosynthesis. However, expression profiling of these genes in the local black soybean varieties is still limited. This study investigated the gene expression levels of the CHS8, CHI1A, IFS2, and CHR genes in local varieties, namely, UP106 (high isoflavone) and UP122 (low isoflavone) and its progenies, i.e., UP106xUP122 and UP122xUP106. Relative gene expression profiling was conducted on the basis of Reverse Transcriptase Polymerase Chain Reaction (RT‐PCR) with ACT2/7 as a housekeeping gene. As a result, the expression level of CHS8 in UP122 is lower than that in UP106. No significant difference in the expression level of CHI1A was observed in all samples. The expression levels of CHS8 and CHI1A in both progenies were higher than that in the parental line, whereas the expression levels of IFS2 in both progenies were lower than that in the parental line. CHS8 and IFS2 expression from UP106xUP122 was higher than that from UP122xUP106, whereas CHI1A expression from UP122xUP106 was higher than that from UP106xUP122. CHR showed a high expression in the reciprocal cross; however, this expression did not exceed from UP106. In conclusion, the crossing between parental lines did not affect the gene expression level in the isoflavone biosynthesis pathway.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"24 1","pages":"119-124"},"PeriodicalIF":0.0,"publicationDate":"2020-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46995000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cloning and in silico study of an endoglucanase from a thermophilic bacterium isolated from a hydrothermal vent of West Kawio, Sangihe‐Talaud waters, North Sulawesi, Indonesia 从印度尼西亚北苏拉威西Sangihe - Talaud水域West Kawio热液喷口分离的嗜热细菌中分离的内切葡聚糖酶的克隆和计算机研究

Indonesian Journal of Biotechnology Pub Date : 2020-01-02 DOI: 10.22146/ijbiotech.48272

Edvan Arifsaputra Suherman, M. Moeis, E. Restiawaty

{"title":"Cloning and in silico study of an endoglucanase from a thermophilic bacterium isolated from a hydrothermal vent of West Kawio, Sangihe‐Talaud waters, North Sulawesi, Indonesia","authors":"Edvan Arifsaputra Suherman, M. Moeis, E. Restiawaty","doi":"10.22146/ijbiotech.48272","DOIUrl":"https://doi.org/10.22146/ijbiotech.48272","url":null,"abstract":"Endoglucanase is used in industries that apply high temperatures, such as bioethanol, detergent, paper, and animal feed industries. Most available endoglucanases have very low stability at high temperatures. Therefore, this study aimed to identfy a new thermostable endoglucanase that is able to maintain its actvity at high temperatures. Five isolates of thermophilic bacteria were previously isolated from the hydrothermal vent of West Kawio, Indonesia. Among them, the DSI2 isolate showed the highest endoglucanase actvity, and was identfed and named as Bacillus safensis DSI2. The Eg DSI2 gene was cloned from B. safensis DSI2. Eg DSI2 is 1851 bp long encoding a protein of 616 amino acids. The encoded protein, EgDSI2, has high sequence identty to other B. safensis endoglucanases and was predicted with the Compute pI/Mw tool to be 69.41 kDa. EgDSI2 was high in hydrophobic amino acids. The enzyme had higher percentage of Ala and Pro, and lower percentage of Gly compared to thermolabile endoglucanases from two Bacillus species. EgDSI2 harbored a catalytc domain belonging to glycosyl hydrolase family 9 (GH9) and a type 3 cellulose‐binding domain (CBM3). Propertes of endoglucanases with GH9‐CBM3 modular organizaton include actvity over a wide pH range, high optmum temperature, and thermostablity. Therefore, EgDSI2 has potental applicatons in the industries.","PeriodicalId":13452,"journal":{"name":"Indonesian Journal of Biotechnology","volume":"24 1","pages":"106-118"},"PeriodicalIF":0.0,"publicationDate":"2020-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46770724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2