G. L. Utama, Mutiara Nabila, H. R. Arifin, E. Lembong, T. Rialita
The research aimed to identify indigenous yeast antibacterial activity from sapodilla fruit against Escherichia coli and Staphylococcus aureus, which conducted by experimental methods and followed by descriptive analysis. This study was done by the isolation of indigenous yeast, macroscopic and microscopic identification, yeast identification using RapID Yeast Plus System, antibacterial test by measuring the clear zone diameter, testing of pathogenic bacteria viability against indigenous yeast and identification of organic acid produced by yeast. The results of yeast isolation obtained 1 isolate (Saccharomyces cereviseae 1) from fruit and 3 isolates form sapodilla skin (S.cereviseae 2, Candida famata, and Pichia anomala) which had antibacterial activity against E. coli and S. aureus except C. famata isolates. Isolates with the largest antibacterial activity against E. coli and S. aureus based on the clear zone diameter were S. cerevisiae (2) isolates. The results of organic acid analysis by HPLC found that S.cerevisiae (2) isolate produced the highest organic acid namely acetic acid as much as 2.442 mg mL -1.
本研究旨在鉴定皂角果对大肠杆菌和金黄色葡萄球菌的抑菌活性，采用实验方法并进行描述性分析。本研究通过对本地酵母的分离、宏观和微观鉴定、快速酵母Plus系统对酵母的鉴定、测定清带直径的抑菌试验、对本地酵母病原菌活力的测定以及酵母产生的有机酸的鉴定等方面进行了研究。酵母菌分离结果表明，除葡萄球菌外，从果实中分离出1株(酿酒酵母菌1)和从果树皮中分离出3株(酿酒酵母菌2、fam念珠菌和异常毕赤酵母)对大肠杆菌和金黄色葡萄球菌均有抑菌活性。从透明带直径来看，对大肠杆菌和金黄色葡萄球菌抑菌活性最大的菌株是酿酒葡萄球菌(2)。HPLC分析结果显示，酿酒酵母(2)分离物的有机酸含量最高，为乙酸2.442 mg mL -1。
{"title":"Antibacterial Activity Test of Indigenous Yeast from Sapodilla Fruit against Staphylococcus aureus and Escherichia coli","authors":"G. L. Utama, Mutiara Nabila, H. R. Arifin, E. Lembong, T. Rialita","doi":"10.5454/mi.13.4.1","DOIUrl":"https://doi.org/10.5454/mi.13.4.1","url":null,"abstract":"The research aimed to identify indigenous yeast antibacterial activity from sapodilla fruit against Escherichia coli and Staphylococcus aureus, which conducted by experimental methods and followed by descriptive analysis. This study was done by the isolation of indigenous yeast, macroscopic and microscopic identification, yeast identification using RapID Yeast Plus System, antibacterial test by measuring the clear zone diameter, testing of pathogenic bacteria viability against indigenous yeast and identification of organic acid produced by yeast. The results of yeast isolation obtained 1 isolate (Saccharomyces cereviseae 1) from fruit and 3 isolates form sapodilla skin (S.cereviseae 2, Candida famata, and Pichia anomala) which had antibacterial activity against E. coli and S. aureus except C. famata isolates. Isolates with the largest antibacterial activity against E. coli and S. aureus based on the clear zone diameter were S. cerevisiae (2) isolates. The results of organic acid analysis by HPLC found that S.cerevisiae (2) isolate produced the highest organic acid namely acetic acid as much as 2.442 mg mL -1.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82420259","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}
Abstract. Tempeh is the most famous traditional fermented food in Indonesia. Tempeh fermentation consists of two stages. In the first stage, the acidification of soybeans used bacteria around 24 hours. Lactic acid bacteria are found in tempeh. Therefore, this study is aimed to investigate the diversity of LAB from tempeh based on 16S rRNA gene sequences and to study their function in tempeh fermentation. In this study, twenty-two LAB isolates were obtained from tempeh. The isolates were closely related to Lactobacillus agilis, Lactobacillus fermentum, Weissella confusa, and Lactobacillus delbrueckii. L. fermentum (13 isolates) were the most abundant in tempeh, followed by L. agilis (7 isolates). It was found LAB important for the acidification of soybeans which the pH of soybean soaking water decreased from pH 7 to pH 4.4-4.9. Key words: diversity,LAB, Lactobacillus,tempeh, Weissella
{"title":"Lactic Acid Bacteria from Tempeh and Their Ability to Acidify Soybeans in Tempeh Fermentation","authors":"Tati Barus, Gabriela Giovania, B. Lay","doi":"10.5454/MI.14.4.4","DOIUrl":"https://doi.org/10.5454/MI.14.4.4","url":null,"abstract":"Abstract. Tempeh is the most famous traditional fermented food in Indonesia. Tempeh fermentation consists of two stages. In the first stage, the acidification of soybeans used bacteria around 24 hours. Lactic acid bacteria are found in tempeh. Therefore, this study is aimed to investigate the diversity of LAB from tempeh based on 16S rRNA gene sequences and to study their function in tempeh fermentation. In this study, twenty-two LAB isolates were obtained from tempeh. The isolates were closely related to Lactobacillus agilis, Lactobacillus fermentum, Weissella confusa, and Lactobacillus delbrueckii. L. fermentum (13 isolates) were the most abundant in tempeh, followed by L. agilis (7 isolates). It was found LAB important for the acidification of soybeans which the pH of soybean soaking water decreased from pH 7 to pH 4.4-4.9. Key words: diversity,LAB, Lactobacillus,tempeh, Weissella","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"46 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89563449","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}
Dengue infection is a global infectious disease with almost 100 million cases occur annually in over more than 100 endemic countries. Dengue virus (DENV), the causative agent of dengue infection, is an 11 kbp RNApositive strand virus which encode 3 structural and 7 non-structural protein within its genome. Non-structural 1 (NS1) protein of DENV is expressed in the earlier stage of infection and having pathogenic role in disease severity. NS1 gene of DENV serotype-2 Indonesian strain was amplified through PCR method using specific designated primers. NS1 amplicon were then cloned into pUMVC4.a and pcDNA3.1 mammalian expression vector which confirmed through colony PCR and sequencing method. Recombinant pUNS1 and pcNS1 plasmids were transfected into CHO-K1 mammalian cell line with lipid based method. Recombinant NS1 protein expression were analyzed through immunostaining using dengue patient sera and rapid NS1 detection kit. Recombinant pUNS1 and pcNS1 plasmids were successfully constructed and recombinant NS1 protein was expressed in CHOK1 mammalian cell line and shown to be reactive against dengue patient sera. Our recombinant NS1 protein also tend to be released outside the transfected CHO-K1 cells as detected in rapid NS1 detection kit. Recombinant dengue NS1 protein was expressed in mammalian cell line in both intra and extracellularly and shown to be immunogenic.
{"title":"Expression of Recombinant Non Structural 1 Protein of Dengue Virus Serotype-2 in Mammalian Cell Line","authors":"I. Rusmana","doi":"10.5454/MI.13.1.%P","DOIUrl":"https://doi.org/10.5454/MI.13.1.%P","url":null,"abstract":"Dengue infection is a global infectious disease with almost 100 million cases occur annually in over more than 100 endemic countries. Dengue virus (DENV), the causative agent of dengue infection, is an 11 kbp RNApositive strand virus which encode 3 structural and 7 non-structural protein within its genome. Non-structural 1 (NS1) protein of DENV is expressed in the earlier stage of infection and having pathogenic role in disease severity. NS1 gene of DENV serotype-2 Indonesian strain was amplified through PCR method using specific designated primers. NS1 amplicon were then cloned into pUMVC4.a and pcDNA3.1 mammalian expression vector which confirmed through colony PCR and sequencing method. Recombinant pUNS1 and pcNS1 plasmids were transfected into CHO-K1 mammalian cell line with lipid based method. Recombinant NS1 protein expression were analyzed through immunostaining using dengue patient sera and rapid NS1 detection kit. Recombinant pUNS1 and pcNS1 plasmids were successfully constructed and recombinant NS1 protein was expressed in CHOK1 mammalian cell line and shown to be reactive against dengue patient sera. Our recombinant NS1 protein also tend to be released outside the transfected CHO-K1 cells as detected in rapid NS1 detection kit. Recombinant dengue NS1 protein was expressed in mammalian cell line in both intra and extracellularly and shown to be immunogenic.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"72 1","pages":"36-42"},"PeriodicalIF":0.0,"publicationDate":"2019-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73292375","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}
O. N. Turnip, Rahma F. Hayati, Rizka Alawiyah, B. Yohan, Dionisius Denis, A. Bowolaksono, A. Soebandrio, R. T. Sasmono
Chikungunya (CHIK) fever, a febrile illness caused by Chikungunya virus (CHIKV) infection, is one of mosquito-borne viral diseases affecting people living in the tropical and subtropical regions in the world. The pathogenesis of the disease is yet to be completely unraveled, and research on CHIK has been conducted by employing various methods, including using cell lines to investigate the biological characteristics of CHIKV in vitro. To assess the suitability of human cell line model for CHIK study, various human cell lines including A549, Huh7, and HepG2 were infected with CHIKV and assayed for their susceptibility to infection. The MTT and plaque assay methods were performed to measure cell viability and virus growth kinetics, respectively. Fluorescence-activated Cell Sorting (FACS) and immunofluorescence assay were performed to measure the proportion of infected cells in the system and their morphological visualization. Both A549 and Huh7 human cell lines showed stable high cell viability upon infection while CHIKV growth kinetics were significantly lower in these cells compared to Vero-CCL81, a monkey cell line that is routinely used in other arboviruses research. Interestingly, we observed significantly different results in HepG2 human cell line, in which cell viability and CHIKV growth kinetics were significantly higher. FACS and immunofluorescence assay confirm the higher infection rate of CHIKV in HepG2 than A549 human cell line. We concluded herethat human hepatocytes HepG2 cell line was susceptible to Asian Genotype of CHIKV and proposed as an alternative cell for the in vitro CHIKV studies to the commonly used A549 and Vero cells.
{"title":"Growth Characteristics of Chikungunya Virus Isolate from Indonesia in Various Human Cell Lines in vitro","authors":"O. N. Turnip, Rahma F. Hayati, Rizka Alawiyah, B. Yohan, Dionisius Denis, A. Bowolaksono, A. Soebandrio, R. T. Sasmono","doi":"10.5454/mi.13.1.1","DOIUrl":"https://doi.org/10.5454/mi.13.1.1","url":null,"abstract":"Chikungunya (CHIK) fever, a febrile illness caused by Chikungunya virus (CHIKV) infection, is one of mosquito-borne viral diseases affecting people living in the tropical and subtropical regions in the world. The pathogenesis of the disease is yet to be completely unraveled, and research on CHIK has been conducted by employing various methods, including using cell lines to investigate the biological characteristics of CHIKV in vitro. To assess the suitability of human cell line model for CHIK study, various human cell lines including A549, Huh7, and HepG2 were infected with CHIKV and assayed for their susceptibility to infection. The MTT and plaque assay methods were performed to measure cell viability and virus growth kinetics, respectively. Fluorescence-activated Cell Sorting (FACS) and immunofluorescence assay were performed to measure the proportion of infected cells in the system and their morphological visualization. Both A549 and Huh7 human cell lines showed stable high cell viability upon infection while CHIKV growth kinetics were significantly lower in these cells compared to Vero-CCL81, a monkey cell line that is routinely used in other arboviruses research. Interestingly, we observed significantly different results in HepG2 human cell line, in which cell viability and CHIKV growth kinetics were significantly higher. FACS and immunofluorescence assay confirm the higher infection rate of CHIKV in HepG2 than A549 human cell line. We concluded herethat human hepatocytes HepG2 cell line was susceptible to Asian Genotype of CHIKV and proposed as an alternative cell for the in vitro CHIKV studies to the commonly used A549 and Vero cells.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90828007","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}
Lipase is a biocatalyst widely used in industry, for example detergent, pharmaceutical, food, or oil purification. One of the most widely lipase used for oil purification is lysophospholipase. As much as 50% of industrial enzyme needs are supplied from microorganisms. However, enzyme productivity from wild type microbial strain is usually limited and not applicable in industry, so that genetic engineering is necessary. Cloning gene encoding for lysophospholipase from Aspergillus niger and Cryptococcus neoformans have been conducted, but has never been conducted from alkalothermophilic bacteria, such as Bacillus halodurans. Bacillus halodurans CM1 is an alkalothermophilic bacterial strain isolated previously that has many industrially potential enzymes. This study aimed to isolate one of the gene encoding lipase from Bacillus halodurans CM1 and cloned into Escherichia coli DH5α using the pGEM-T easy vector. The gene fragment encoding lysophospholipase obtained with size 783 base pairs and had 100% similarity with gene encoding lysophospholipase from Bacillus halodurans C-125 (No access GenBank: BA000004.3). E. coli harbouring the recombinant plasmid with the gene also showed activity on trybutiryn medium compared to negative control.
脂肪酶是一种广泛应用于洗涤剂、制药、食品、油脂净化等工业领域的生物催化剂。溶血磷脂酶是用于油脂净化的最广泛的脂肪酶之一。多达50%的工业酶需求是由微生物提供的。然而，野生型微生物菌株的酶产量通常是有限的，不能应用于工业，因此基因工程是必要的。从黑曲霉和新生隐球菌中克隆溶血磷脂酶编码基因已经进行过，但从未从嗜碱热细菌中克隆过，如嗜盐芽孢杆菌。嗜盐芽孢杆菌(Bacillus halodurans) CM1是一种嗜碱热细菌菌株，具有许多工业上潜在的酶。本研究旨在从嗜盐芽孢杆菌CM1中分离一个脂肪酶编码基因，并利用pgm - t easy载体将其克隆到大肠杆菌DH5α中。编码溶血磷脂酶的基因片段大小为783个碱基对，与嗜盐芽孢杆菌C-125(无访问GenBank: BA000004.3)中编码溶血磷脂酶的基因具有100%的相似性。与阴性对照相比，携带该基因的重组质粒的大肠杆菌在trybutyrin培养基上也表现出活性。
{"title":"Isolation of a Functional Gene Encoding Homologous Lysophospholipase from Indonesian Indigenous Bacillus halodurans CM1","authors":"S. Fernanda, A. Abinawanto, I. Helianti","doi":"10.5454/mi.13.1.2","DOIUrl":"https://doi.org/10.5454/mi.13.1.2","url":null,"abstract":"Lipase is a biocatalyst widely used in industry, for example detergent, pharmaceutical, food, or oil purification. One of the most widely lipase used for oil purification is lysophospholipase. As much as 50% of industrial enzyme needs are supplied from microorganisms. However, enzyme productivity from wild type microbial strain is usually limited and not applicable in industry, so that genetic engineering is necessary. Cloning gene encoding for lysophospholipase from Aspergillus niger and Cryptococcus neoformans have been conducted, but has never been conducted from alkalothermophilic bacteria, such as Bacillus halodurans. Bacillus halodurans CM1 is an alkalothermophilic bacterial strain isolated previously that has many industrially potential enzymes. This study aimed to isolate one of the gene encoding lipase from Bacillus halodurans CM1 and cloned into Escherichia coli DH5α using the pGEM-T easy vector. The gene fragment encoding lysophospholipase obtained with size 783 base pairs and had 100% similarity with gene encoding lysophospholipase from Bacillus halodurans C-125 (No access GenBank: BA000004.3). E. coli harbouring the recombinant plasmid with the gene also showed activity on trybutiryn medium compared to negative control.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"43 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89157260","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}
Identification of endophytic fungi in Paraserianthes falcatria is the effort of the potential of endophytic fungi as phytohormone producer. Phytohormone is needed to spur shoot and root initiation. This study in P. falcatariais necessary when woody of P. falcataria decreases every year. The aimed of the study were to identify endophytic fungi from leaves, twigs, and roots of P. falcataria, and determine IAA content from endophytic fungi. Isolates that were grown from leaves, twigs and roots cuttings on PDA, were identified based on micro- and macromorphology. Determining of IAA content was counted with spectrophotometer vis based on a calibration curve from the standard solution. The results were obtained 10 of isolates fungi from leaves, twigs, and roots. But from 10, only nine isolates that could be identified. They were Aspergillus sp., Acremonium sp., Cladosporium sp., Trichoderma sp. 1, Phytium sp., Rhizoctonia sp., Trichoderma sp. 2, Hormiscium sp. 1 and Hormiscium sp. 2. Production of indole acetic acid (IAA) from Cladosporium sp. had the highest content than others (311 ppm). The lowest IAA content (51.97 ppm) was produced by the Rhizoctonia sp. The study can be continued to find out their abilities as PGPF agents and biopesticides of P. falcataria seedlings.
{"title":"Endophytic Fungi in Paraserianthes falcataria: Production of Indole Acetic Acid","authors":"R. S. Wulandari, R. Suryantini","doi":"10.5454/mi.13.1.3","DOIUrl":"https://doi.org/10.5454/mi.13.1.3","url":null,"abstract":"Identification of endophytic fungi in Paraserianthes falcatria is the effort of the potential of endophytic fungi as phytohormone producer. Phytohormone is needed to spur shoot and root initiation. This study in P. falcatariais necessary when woody of P. falcataria decreases every year. The aimed of the study were to identify endophytic fungi from leaves, twigs, and roots of P. falcataria, and determine IAA content from endophytic fungi. Isolates that were grown from leaves, twigs and roots cuttings on PDA, were identified based on micro- and macromorphology. Determining of IAA content was counted with spectrophotometer vis based on a calibration curve from the standard solution. The results were obtained 10 of isolates fungi from leaves, twigs, and roots. But from 10, only nine isolates that could be identified. They were Aspergillus sp., Acremonium sp., Cladosporium sp., Trichoderma sp. 1, Phytium sp., Rhizoctonia sp., Trichoderma sp. 2, Hormiscium sp. 1 and Hormiscium sp. 2. Production of indole acetic acid (IAA) from Cladosporium sp. had the highest content than others (311 ppm). The lowest IAA content (51.97 ppm) was produced by the Rhizoctonia sp. The study can be continued to find out their abilities as PGPF agents and biopesticides of P. falcataria seedlings.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"16 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83146491","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}
Our previous research had screened 9 best indigenous endophytic isolates for their ability to control Ralstonia syzigii subsp. indonesiensis, the causal agents of bacterial wilt disease in tomato ( Lycopersicon esculentum ) in green house condition. Those 9 strains were B acillus cereus EPL1.1.3 , B. cereus TLE2.3, B. toyonensis EPL1.1.4 , Serratia nematodiphila TLE1.1, B. anthracis SNE2.2, B. cereus E1.AB1.2, B. cereus E1AB2.1, Enterobacter cloacae subsp. dissolvens TLE2.2 and S. marcescens KLE3.3. The purposed of this study is to test the ability of the endophytic bacteria strains in increasing defense related enzyme activities of tomato. Bacterial strains were tested for its ability to induce the defense-related enzymes which were phenylalanine ammonia lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) in roots and leaves of tomato plants. R. syzigii subsp. indonesiensis inoculated to host plants 7 days after the endophyte bacteria strains inoculation. Enzyme activities were recorded at 0, 1, 3, 5, 7, 9, 12 and 15 days after pathogen inoculation (dpi). It was observed that PAL, PO and PPO activities were significantly increased in all of the endophytic bacteria inoculated treatments compared to control plant. Activities of PAL in the leaves was fast similar to the roots; but PO activities was higher in the roots compared to that in the leaves, whereas PPO activities was higher in the leaves than in the roots. PAL and PO reached the maximum level at different time in the leaves (3 dpi and 15 dpi), in the roots (5 dpi and 12 dpi), whereas PPO in the leaves at 12 dpi and in the roots at 9 dpi.
{"title":"Induced Defense Related Enzyme Activities of Tomato Plant by Indigenous Endophytic Bacteria and Challenged by Ralstonia Syzigii Subsp. Indonesiensis","authors":"Y. Yanti, W. Warnita, R. Reflin","doi":"10.5454/MI.13.1.4","DOIUrl":"https://doi.org/10.5454/MI.13.1.4","url":null,"abstract":"Our previous research had screened 9 best indigenous endophytic isolates for their ability to control Ralstonia syzigii subsp. indonesiensis, the causal agents of bacterial wilt disease in tomato ( Lycopersicon esculentum ) in green house condition. Those 9 strains were B acillus cereus EPL1.1.3 , B. cereus TLE2.3, B. toyonensis EPL1.1.4 , Serratia nematodiphila TLE1.1, B. anthracis SNE2.2, B. cereus E1.AB1.2, B. cereus E1AB2.1, Enterobacter cloacae subsp. dissolvens TLE2.2 and S. marcescens KLE3.3. The purposed of this study is to test the ability of the endophytic bacteria strains in increasing defense related enzyme activities of tomato. Bacterial strains were tested for its ability to induce the defense-related enzymes which were phenylalanine ammonia lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) in roots and leaves of tomato plants. R. syzigii subsp. indonesiensis inoculated to host plants 7 days after the endophyte bacteria strains inoculation. Enzyme activities were recorded at 0, 1, 3, 5, 7, 9, 12 and 15 days after pathogen inoculation (dpi). It was observed that PAL, PO and PPO activities were significantly increased in all of the endophytic bacteria inoculated treatments compared to control plant. Activities of PAL in the leaves was fast similar to the roots; but PO activities was higher in the roots compared to that in the leaves, whereas PPO activities was higher in the leaves than in the roots. PAL and PO reached the maximum level at different time in the leaves (3 dpi and 15 dpi), in the roots (5 dpi and 12 dpi), whereas PPO in the leaves at 12 dpi and in the roots at 9 dpi.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81037547","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}
V. Kurnianda, S. Faradilla, S. Karina, S. Agustina, M. Ulfah, C. Octavina, F. Syahliza, M. R. Ramadhan, S. Purnawan, M. Musman
Dengue infection is a global health problem with an increasing incidence every year and now endemic in more than 100 WHO countries. Dengue infection is caused by dengue virus (DENV) which is an RNA virus with positive single strand, with ±11kb genome size encoding 3 structural proteins, 7 non-structural proteins, and two Untranslated Region (UTR). NS1 protein is known to have a very important role in the development of severe DENV infection, by the direct effect causing host cells damage and indirect effect by activating immune response to induce the secretion of excess cytokines. Transfected CHO-K1 cells with recombinant pcNS1 plasmid was co-cultured with PBMC from healthy donor. 48 hours post co-cultured, cell supernatant was collected and TNF-α levels and NS1 recombinant were measured by ELISA. Recombinant NS1 protein was expressed in CHO-K1 mammalian cell line and able to induce TNF-α with higher levels compared to control. Recombinant NS1 protein expression was able to induce secretion of TNF-α with higher levels compared to control.
{"title":"Levels of TNF-α in PBMC (Peripheral Blood Mononuclear Cells) Induced by Recombinant Non Structural 1 Protein of Dengue Virus Serotype-2 in vitro","authors":"V. Kurnianda, S. Faradilla, S. Karina, S. Agustina, M. Ulfah, C. Octavina, F. Syahliza, M. R. Ramadhan, S. Purnawan, M. Musman","doi":"10.5454/mi.13.2.%p","DOIUrl":"https://doi.org/10.5454/mi.13.2.%p","url":null,"abstract":"Dengue infection is a global health problem with an increasing incidence every year and now endemic in more than 100 WHO countries. Dengue infection is caused by dengue virus (DENV) which is an RNA virus with positive single strand, with ±11kb genome size encoding 3 structural proteins, 7 non-structural proteins, and two Untranslated Region (UTR). NS1 protein is known to have a very important role in the development of severe DENV infection, by the direct effect causing host cells damage and indirect effect by activating immune response to induce the secretion of excess cytokines. Transfected CHO-K1 cells with recombinant pcNS1 plasmid was co-cultured with PBMC from healthy donor. 48 hours post co-cultured, cell supernatant was collected and TNF-α levels and NS1 recombinant were measured by ELISA. Recombinant NS1 protein was expressed in CHO-K1 mammalian cell line and able to induce TNF-α with higher levels compared to control. Recombinant NS1 protein expression was able to induce secretion of TNF-α with higher levels compared to control.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"55 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74035179","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}
Grace DOLOROSA LIMBONG, Levy NATHANAEL NABABAN, A. Manurung, Merry MERYAM MARTGRITA
According to several studies, solid state fermentation (SSF) can enhance antioxidant and antibacterial activity of natural sources, and microorganism that is widely used in this kind of research is Aspergillus oryzae . Therefore, this study employed SSF by A. oryzae to enhance antioxidant and antibacterial activity of candlenut kernel. Candlenut kernel powder, that has been moistened with 60% water, was inoculated with 10% (w/w) of 5-day-culture of A. oryzae, and was fermented for 9 days (until exponential phase; sample-1) and 12 days (until stationary phase; sample-2). The fermented candlenut kernels was extracted by ethanol and concentrated using rotary evaporator. Total phenolic content of control (unfermented extract), sample-1, and sample-2 are 0.183 mg -1 -1 -1 GAE g , 2.761 mg GAE g , and 4.194 mg GAE g , respectively. This results supported the IC value determined 50 -1 -1 -1 by DPPH (2,2-diphenyl-1-picrylhydrazyl) method, those are 617.11 μ g mL , 260.23 μ g mL , and 45.29 μ g mL . -1 These results revealed a very strong antioxidant activity (< 50 μ g mL ) in the sample fermented until stationary phase. Antibacterial assay against Staphylococcus aureus resulted diameter of inhibition zone 7.17 mm, 13.51 mm, and 18.51 mm, respectively; whereas against Pseudomonas aeruginosa resulted diameter of inhibition zone 6.52 mm, 11.786 mm, and 15.269 mm, respectively. From this result, SSF until stationary phase enhanced higher antioxidant and antibacterial activity compared the other treatments.
根据多项研究，固态发酵(SSF)可以增强天然来源的抗氧化和抗菌活性，而在这类研究中被广泛使用的微生物是米曲霉。因此，本研究利用A. oryzae的SSF增强了核桃仁的抗氧化和抗菌活性。用60%的水润湿的核桃仁粉，接种10% (w/w)的5天培养量的稻瘟病菌，发酵9天(至指数期;样品-1)和12天(直到固定阶段;示例2)。用乙醇提取发酵后的香烛仁，用旋转蒸发器浓缩。对照(未发酵提取物)、样品-1和样品-2的总酚含量分别为0.183 mg -1 -1 GAE g、2.761 mg GAE g和4.194 mg GAE g。结果支持DPPH(2,2-二苯基-1-吡啶肼基)法测定的IC值为50 -1 -1 -1，分别为617.11 μ g mL、260.23 μ g mL和45.29 μ g mL。这些结果表明，发酵至固定相的样品具有很强的抗氧化活性(< 50 μ g mL)。对金黄色葡萄球菌的抑菌实验结果显示，抑菌带直径分别为7.17 mm、13.51 mm和18.51 mm;对铜绿假单胞菌的抑菌带直径分别为6.52 mm、11.786 mm和15.269 mm。从这个结果来看，与其他处理相比，SSF在固定阶段具有更高的抗氧化和抗菌活性。
{"title":"Antioxidant and Antibacterial Activities Enhancement of Solid-state Fermented Candlenut Kernels by Aspergillus oryzae","authors":"Grace DOLOROSA LIMBONG, Levy NATHANAEL NABABAN, A. Manurung, Merry MERYAM MARTGRITA","doi":"10.5454/mi.13.2.2","DOIUrl":"https://doi.org/10.5454/mi.13.2.2","url":null,"abstract":"According to several studies, solid state fermentation (SSF) can enhance antioxidant and antibacterial activity of natural sources, and microorganism that is widely used in this kind of research is Aspergillus oryzae . Therefore, this study employed SSF by A. oryzae to enhance antioxidant and antibacterial activity of candlenut kernel. Candlenut kernel powder, that has been moistened with 60% water, was inoculated with 10% (w/w) of 5-day-culture of A. oryzae, and was fermented for 9 days (until exponential phase; sample-1) and 12 days (until stationary phase; sample-2). The fermented candlenut kernels was extracted by ethanol and concentrated using rotary evaporator. Total phenolic content of control (unfermented extract), sample-1, and sample-2 are 0.183 mg -1 -1 -1 GAE g , 2.761 mg GAE g , and 4.194 mg GAE g , respectively. This results supported the IC value determined 50 -1 -1 -1 by DPPH (2,2-diphenyl-1-picrylhydrazyl) method, those are 617.11 μ g mL , 260.23 μ g mL , and 45.29 μ g mL . -1 These results revealed a very strong antioxidant activity (< 50 μ g mL ) in the sample fermented until stationary phase. Antibacterial assay against Staphylococcus aureus resulted diameter of inhibition zone 7.17 mm, 13.51 mm, and 18.51 mm, respectively; whereas against Pseudomonas aeruginosa resulted diameter of inhibition zone 6.52 mm, 11.786 mm, and 15.269 mm, respectively. From this result, SSF until stationary phase enhanced higher antioxidant and antibacterial activity compared the other treatments.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"15 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85066428","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}
Agarwood is a type of semitoleran plant, so that for planting the seedlings should be grown under the shade. For planting in open land, it requires treatment in which one of them is using seedlings inoculated with arbuscularmycorrhizal fungi. The Aim of the research were: (1) to obtain information on agarwood growth that has been inoculated with fungi mycorrhizalarbuscular when planted in the open land, and (2) ability to grow between agarwood seedlings inoculated mycorrhizal that was planted in the shade and in the open area. Split Plot randomized block design was applied with treatments : the first plot consisting of plant had been inoculated with mycorrhizae and without mycorrhizal inoculation, and the sub plot was the types of shading that consists of : open land, paranet 60 % intensity and natural vegetation. To reduce variabilty of site topographical differences were separated as bloks. Variables measured were :plant height (cm), stem diameter (mm), number of leaves and survival percentage of plant. The results show that the height and diameter growth of seedlings innoculated with mycorrhizae were higher than non innoculated. The seedlings innoculated with mycorrhizal fungi were planted in the paranet shading grew better and significantly different compared to the vegetation shading. Seedlings innoculatedmycorrhizal that were planted in open land grew better and significantly different compared to vegetation shading. This study results indicate that planting agarwood in the open land can be done using seedlings inoculated arbuscularmycorrhizal fungi.
{"title":"The Utilization of ArbuscularMycorrhizal Fungi For Planting Agarwood (Aquilariaspp) Seedling In Open Land","authors":"A. Muin, Fachrizal Fachrizal","doi":"10.5454/MI.13.3.%P","DOIUrl":"https://doi.org/10.5454/MI.13.3.%P","url":null,"abstract":"Agarwood is a type of semitoleran plant, so that for planting the seedlings should be grown under the shade. For planting in open land, it requires treatment in which one of them is using seedlings inoculated with arbuscularmycorrhizal fungi. The Aim of the research were: (1) to obtain information on agarwood growth that has been inoculated with fungi mycorrhizalarbuscular when planted in the open land, and (2) ability to grow between agarwood seedlings inoculated mycorrhizal that was planted in the shade and in the open area. Split Plot randomized block design was applied with treatments : the first plot consisting of plant had been inoculated with mycorrhizae and without mycorrhizal inoculation, and the sub plot was the types of shading that consists of : open land, paranet 60 % intensity and natural vegetation. To reduce variabilty of site topographical differences were separated as bloks. Variables measured were :plant height (cm), stem diameter (mm), number of leaves and survival percentage of plant. The results show that the height and diameter growth of seedlings innoculated with mycorrhizae were higher than non innoculated. The seedlings innoculated with mycorrhizal fungi were planted in the paranet shading grew better and significantly different compared to the vegetation shading. Seedlings innoculatedmycorrhizal that were planted in open land grew better and significantly different compared to vegetation shading. This study results indicate that planting agarwood in the open land can be done using seedlings inoculated arbuscularmycorrhizal fungi.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91257955","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}
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