W. Munaeni, A. Pariakan, Laode Baytul Abidin, M. Yuhana
The objectives of this study were to analyze phytochemical content of bawang hutan bulbs extract ( Eleutherine palmifolia ) and to test the inhibitory potential of bawang hutan bulbs extract on the growth of Vibrio harveyi bacteria at different doses. This study was conducted in March-May 2017 in Testing Laboratory of Fisheries and Marine Science Faculty of Halu Oleo University and Laboratory of Fish Health of Aquaculture Department of Fisheries and Marine Science Faculty and Laboratory of Biopharmaca of Bogor Agricultural University. Test parameter included: (1) Phytochemical test through the method of color visualization, (2) Inhibitory potential test using two methods namely agar diffusion and co-culture. Treatment of dose consisted of positive control/K+ (Chloramphenicol 30 mg/ml), negative control/K- (Sterile Aquadest) and treatment of extract included A (20 mg/ml), B (40 mg/ml), C (60 mg/ml), D (80 mg/ml). Qualitatively, result of phytochemical test showed that bawang hutan bulbs extract contained flavonoid, tannin, saponin, quinone, steroid and triterpenoid compounds. Result of inhibitory potential test indicated that treatment D obtained the highest inhibitory potential, while the minimum inhibitory potential was found in treatment A. The best co-culture test result was also found in treatment D, in which 24 hours after co-culture was performed, no V. harveyi colonies (total bacteria of 0 CFU/mL) were found. Bawang hutan bulbs extract in this study was able to inhibit the growth of V. harveyi.
{"title":"In Vitro Phytochemical and Inhibitory Potential Test of Bawang Hutan Bulb Extract (Eleutherine palmifolia) on Vibrio harveyi","authors":"W. Munaeni, A. Pariakan, Laode Baytul Abidin, M. Yuhana","doi":"10.5454/MI.11.3.%P","DOIUrl":"https://doi.org/10.5454/MI.11.3.%P","url":null,"abstract":"The objectives of this study were to analyze phytochemical content of bawang hutan bulbs extract ( Eleutherine palmifolia ) and to test the inhibitory potential of bawang hutan bulbs extract on the growth of Vibrio harveyi bacteria at different doses. This study was conducted in March-May 2017 in Testing Laboratory of Fisheries and Marine Science Faculty of Halu Oleo University and Laboratory of Fish Health of Aquaculture Department of Fisheries and Marine Science Faculty and Laboratory of Biopharmaca of Bogor Agricultural University. Test parameter included: (1) Phytochemical test through the method of color visualization, (2) Inhibitory potential test using two methods namely agar diffusion and co-culture. Treatment of dose consisted of positive control/K+ (Chloramphenicol 30 mg/ml), negative control/K- (Sterile Aquadest) and treatment of extract included A (20 mg/ml), B (40 mg/ml), C (60 mg/ml), D (80 mg/ml). Qualitatively, result of phytochemical test showed that bawang hutan bulbs extract contained flavonoid, tannin, saponin, quinone, steroid and triterpenoid compounds. Result of inhibitory potential test indicated that treatment D obtained the highest inhibitory potential, while the minimum inhibitory potential was found in treatment A. The best co-culture test result was also found in treatment D, in which 24 hours after co-culture was performed, no V. harveyi colonies (total bacteria of 0 CFU/mL) were found. Bawang hutan bulbs extract in this study was able to inhibit the growth of V. harveyi.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"111 3S 1","pages":"4"},"PeriodicalIF":0.0,"publicationDate":"2017-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72583969","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}
In liquid wastes, especially domestic wastewater, many organic substances are mixed causing water quality degradation, one of them is ammonia. Liquid wastes containing ammonia can be treated using an activated sludge system. One of the active sludge that can be used is shrimp pond sediments. This experiment investigated the performance of microbes in shrimp pond sediments with the addition of EM4 in nitrification process for the treatment of wastewater with high ammonia concentration in a 8 L batch reactor capacity. The results show that the addition of shrimp pond sediment as the active sludge can remove high ammonia level almost completely and there is known interaction between time and variation of shrimp pond quantity (p value <0,05) to the decreasing of ammonia level. Efficiency of decreasing the concentration of ammonia up to 100% can be reached on the 15th day in each treatment. The addition of EM4 can shorten the decreasing of ammonia level by 50%. Keywords : Nitrification, Ammonia, Shrimp Pond Sediment, EM4, Activated Sludge
{"title":"Performance Optimization of Microbes from Shrimp Pond Sediment by Adding EM4 In Nitrification Process for the Treatment of Wastewater Containing High Ammonia Concentration","authors":"H. Ambarsari, M. R. Harahap","doi":"10.5454/MI.11.3.4","DOIUrl":"https://doi.org/10.5454/MI.11.3.4","url":null,"abstract":"In liquid wastes, especially domestic wastewater, many organic substances are mixed causing water quality degradation, one of them is ammonia. Liquid wastes containing ammonia can be treated using an activated sludge system. One of the active sludge that can be used is shrimp pond sediments. This experiment investigated the performance of microbes in shrimp pond sediments with the addition of EM4 in nitrification process for the treatment of wastewater with high ammonia concentration in a 8 L batch reactor capacity. The results show that the addition of shrimp pond sediment as the active sludge can remove high ammonia level almost completely and there is known interaction between time and variation of shrimp pond quantity (p value <0,05) to the decreasing of ammonia level. Efficiency of decreasing the concentration of ammonia up to 100% can be reached on the 15th day in each treatment. The addition of EM4 can shorten the decreasing of ammonia level by 50%. Keywords : Nitrification, Ammonia, Shrimp Pond Sediment, EM4, Activated Sludge","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"124 1","pages":"4"},"PeriodicalIF":0.0,"publicationDate":"2017-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76698814","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}
M. Anggiani, I. Helianti, N. Nurhayati, A. Abinawanto
Lipase is one of the most important industrial enzymes, which is widely used in the preparation of food additives, cosmetics, and pharmaceutical industries. In the previous study, we have cloned synthetic Thermomyces lanuginosus lipase gene into Bacillus subtilis and Escherichia coli and resulting low expression for enzyme activity. The aim of this research was to construct the Thermomyces lanuginosus lipase (TLL) gene into Pichia pastoris vector expression with TLL original signal peptide. TLL gene was amplified by PCR and contained original signal peptide and then inserted into pPICZα A between XhoI and XbaI site, and transformed into competent cell E.coli DH5α. From the transformant, two of positive recombinants were analyzed by sequencing analysis. As the result, both of two recombinant have a positive target gene which has lipase gene. The correct plasmid was linearized and then was transformed in Pichia pastoris X-33 by electroporation method. Thermomyces lanuginosus synthetic gene lipase has been successfully integrated into chromosome of P. pastoris X-33, which revealed by clear zones arund the colony on Yeast extract Peptone Dextrose Tributyrin (YPD.TB) plate with zeocin. The Thermomyces lanuginosus lipase had an open reading frame of 916bp encoding TLL of 314 amino acids with theoretical molecular mass of 35 kDa. The recombinant enzyme, Thermomyces lanuginosus lipase had optimal temperature at 80˚C and optimal pH at pH 8.
{"title":"Cloning of Lipase Gene From Thermomyces langinosus into Pichia pastoris with its Original Signal Peptide","authors":"M. Anggiani, I. Helianti, N. Nurhayati, A. Abinawanto","doi":"10.5454/MI.11.2.4","DOIUrl":"https://doi.org/10.5454/MI.11.2.4","url":null,"abstract":"Lipase is one of the most important industrial enzymes, which is widely used in the preparation of food additives, cosmetics, and pharmaceutical industries. In the previous study, we have cloned synthetic Thermomyces lanuginosus lipase gene into Bacillus subtilis and Escherichia coli and resulting low expression for enzyme activity. The aim of this research was to construct the Thermomyces lanuginosus lipase (TLL) gene into Pichia pastoris vector expression with TLL original signal peptide. TLL gene was amplified by PCR and contained original signal peptide and then inserted into pPICZα A between XhoI and XbaI site, and transformed into competent cell E.coli DH5α. From the transformant, two of positive recombinants were analyzed by sequencing analysis. As the result, both of two recombinant have a positive target gene which has lipase gene. The correct plasmid was linearized and then was transformed in Pichia pastoris X-33 by electroporation method. Thermomyces lanuginosus synthetic gene lipase has been successfully integrated into chromosome of P. pastoris X-33, which revealed by clear zones arund the colony on Yeast extract Peptone Dextrose Tributyrin (YPD.TB) plate with zeocin. The Thermomyces lanuginosus lipase had an open reading frame of 916bp encoding TLL of 314 amino acids with theoretical molecular mass of 35 kDa. The recombinant enzyme, Thermomyces lanuginosus lipase had optimal temperature at 80˚C and optimal pH at pH 8.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"16 1","pages":"4"},"PeriodicalIF":0.0,"publicationDate":"2017-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75059894","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}
Lisa Pratama, I. Helianti, A. Suryani, Budiasih Wahyuntari
Lipase catalyses hydrolysis and esterification of lipids. The purpose of this research was to obtain lipase producing indigenous fungi, to identify the selected fungi, to study optimum temperature and pH of the enzyme activity, as well as the enzyme ability in interesterification reaction. The isolates used in the experiment were isolated from tempeh, oncom and BPPT laboratory culture collection. The results showed that three fungal isolates which isolated, tempe and oncom and an isolate of BPPT-CC were positive produced lipase after qualitative assay using Rhodamine B, olive oil and PVA. The morphology identification of the isolates, revealed that R isolate was Aspergillus sp, T isolate was Neurospora sp. and O isolate was Rhizopus sp. Upon quantitative assay from determination of the media and time production, potato dextro broth (PDB) with olive oil 2% in 48 hours fermentation showed the highest specific activity of the enzymes. Lipase produced from three isolate have the optimum at pH 4, temperatures at 40-45 °C and stable in interesterification reaction (55 °C) for 30-40 min. HPLC analysis after interesterification enzymatic reaction in mixture palm kernel olein (PKOo) and palm stearin (POs) showed that the composition of triglycerides (TAG) do not change if compared with the commercial lipase (Lypozyme TL1M).
{"title":"Isolation, Characterization, and Production of Lipase from Indigenous Fungal for Enzymatic Interesterification Process","authors":"Lisa Pratama, I. Helianti, A. Suryani, Budiasih Wahyuntari","doi":"10.5454/MI.11.2.1","DOIUrl":"https://doi.org/10.5454/MI.11.2.1","url":null,"abstract":"Lipase catalyses hydrolysis and esterification of lipids. The purpose of this research was to obtain lipase producing indigenous fungi, to identify the selected fungi, to study optimum temperature and pH of the enzyme activity, as well as the enzyme ability in interesterification reaction. The isolates used in the experiment were isolated from tempeh, oncom and BPPT laboratory culture collection. The results showed that three fungal isolates which isolated, tempe and oncom and an isolate of BPPT-CC were positive produced lipase after qualitative assay using Rhodamine B, olive oil and PVA. The morphology identification of the isolates, revealed that R isolate was Aspergillus sp, T isolate was Neurospora sp. and O isolate was Rhizopus sp. Upon quantitative assay from determination of the media and time production, potato dextro broth (PDB) with olive oil 2% in 48 hours fermentation showed the highest specific activity of the enzymes. Lipase produced from three isolate have the optimum at pH 4, temperatures at 40-45 °C and stable in interesterification reaction (55 °C) for 30-40 min. HPLC analysis after interesterification enzymatic reaction in mixture palm kernel olein (PKOo) and palm stearin (POs) showed that the composition of triglycerides (TAG) do not change if compared with the commercial lipase (Lypozyme TL1M).","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"10 1","pages":"1"},"PeriodicalIF":0.0,"publicationDate":"2017-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81857949","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}
Aeromonas hydrophila is a type of bacteria causing Motile Aeromonads Septicemia (MAS) disease, which infects freshwater fish, including African catfish, and leads to death as well as huge losses to farmers. This research aims to determine the resistance status of various antibiotic classes in A. hydrophila bacteria isolated from African catfish. Bacterial isolates of A. hydrophila were taken from the liver and kidneys of infected African catfishes obtained from Parung, Bogor. Characterization of the bacteria was carried out based on colony morphology and biochemical properties. Meanwhile, bacterial resistance test was conducted using antibiotic disks with Kirby-Bauer method. Based on colony morphology and biochemical properties, the characterization results indicated that the bacterial isolates tested were A. hydrophila . Further examination of the antibiotic resistance test showed that the bacteria were resistant to penicillin antibiotics and macrolides. Future researches are expected to use molecular identification for A.hydrophila bacteria mutant to known the DNA base.
{"title":"Resistance Test on Aeromonas hydrophila Isolated from African Catfish (Clarias gariepinus) Against Some Antibiotics Groups","authors":"M. F. Ulkhaq, A. Lusiastuti","doi":"10.5454/MI.11.2.5","DOIUrl":"https://doi.org/10.5454/MI.11.2.5","url":null,"abstract":"Aeromonas hydrophila is a type of bacteria causing Motile Aeromonads Septicemia (MAS) disease, which infects freshwater fish, including African catfish, and leads to death as well as huge losses to farmers. This research aims to determine the resistance status of various antibiotic classes in A. hydrophila bacteria isolated from African catfish. Bacterial isolates of A. hydrophila were taken from the liver and kidneys of infected African catfishes obtained from Parung, Bogor. Characterization of the bacteria was carried out based on colony morphology and biochemical properties. Meanwhile, bacterial resistance test was conducted using antibiotic disks with Kirby-Bauer method. Based on colony morphology and biochemical properties, the characterization results indicated that the bacterial isolates tested were A. hydrophila . Further examination of the antibiotic resistance test showed that the bacteria were resistant to penicillin antibiotics and macrolides. Future researches are expected to use molecular identification for A.hydrophila bacteria mutant to known the DNA base.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"10 1","pages":"5"},"PeriodicalIF":0.0,"publicationDate":"2017-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85139439","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}
C. N. Sari, Usman Usman, Riesa K. W. Rohmat, L. Herlina, Ken Sawitri Suliandri, Onie Kristiawan, Dwiyantari Dwiyantari, T. Kristianti, S. Suhandono
Surfactant is generally synthetic chemical, which is effective and reliable. However, the chemicals usually did not degraded easily in the environment and could cause damage to the environment. The other possible alternative to produce surfactant is using genetic engineering in order to produce peptide based surfactant. In this research, peptide surfactant was produced using a gene construct which was created using overlapped polymerase chain reaction method (OE-PCR). PAGE analysis shows that single surfactant peptide construction can be expressed by induction of IPTG 1 mM and after at least twice sonication. This research proves that both two constructions have been successfully expressed by producing peptide in expected size (approximately 15 kDa).
{"title":"Construction and Expression of Single Recombinant Peptide Surfactant for EOR Application","authors":"C. N. Sari, Usman Usman, Riesa K. W. Rohmat, L. Herlina, Ken Sawitri Suliandri, Onie Kristiawan, Dwiyantari Dwiyantari, T. Kristianti, S. Suhandono","doi":"10.5454/MI.11.1.5","DOIUrl":"https://doi.org/10.5454/MI.11.1.5","url":null,"abstract":"Surfactant is generally synthetic chemical, which is effective and reliable. However, the chemicals usually did not degraded easily in the environment and could cause damage to the environment. The other possible alternative to produce surfactant is using genetic engineering in order to produce peptide based surfactant. In this research, peptide surfactant was produced using a gene construct which was created using overlapped polymerase chain reaction method (OE-PCR). PAGE analysis shows that single surfactant peptide construction can be expressed by induction of IPTG 1 mM and after at least twice sonication. This research proves that both two constructions have been successfully expressed by producing peptide in expected size (approximately 15 kDa).","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"1190 1","pages":"5-5"},"PeriodicalIF":0.0,"publicationDate":"2017-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72742373","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}
M. M. Nuryady, Sugeng Seyo Utomo, Yunita Armiyanti, Sri Mumpuni Wahyu Widjajati, K. Senjarini
Malaria is an infectious disease caused by Plasmodium, which is transmitted by Anopheles mosquitoes as vectors. Malaria transmission begins when an infected mosquito takes blood meal from healthy human. Mosquitoes will release parasite and components of saliva into the host's body. Saliva contains components (proteins) that affect the host's hemostasis and immune respose, such as vasomodulator and immunomodulators. Imunomudulator could act as immunosuppressive factors that can suppress nonspecific immune system of the host and modulate the change of T helper 1 (Th1) toward T helper 2 (Th2) response, which is advantageous for malaria parasite to infect human host. This research wanted to evaluate human immune respons in endemic area against salivary gland protein extract (SGPE) from its major malaria vector i.e. Anopheles sundaicus (An. sundaicus). Analysis of human immune response was conducted quantitatively by ELISA (Enzyme Link Immunosorbend Assay) towards IgG from human sera samples after cross reacted with SGPE. The results showed that exposures to An. sundaicus were able to induce high levels of IgG. IgG anti salivary proteins of An. sundaicus is higher than the levels of IgG anti salivary proteins of Ae. aegypti. Furthermore, the age group 11-40 years with the highest bites probability, had the highest IgG levels compared to other age groups.
{"title":"Analysis of Human Immune Response against Salivary Glands Protein Extract of Anopheles sundaicus. L in Malaria Endemic Area","authors":"M. M. Nuryady, Sugeng Seyo Utomo, Yunita Armiyanti, Sri Mumpuni Wahyu Widjajati, K. Senjarini","doi":"10.5454/MI.11.1.4","DOIUrl":"https://doi.org/10.5454/MI.11.1.4","url":null,"abstract":"Malaria is an infectious disease caused by Plasmodium, which is transmitted by Anopheles mosquitoes as vectors. Malaria transmission begins when an infected mosquito takes blood meal from healthy human. Mosquitoes will release parasite and components of saliva into the host's body. Saliva contains components (proteins) that affect the host's hemostasis and immune respose, such as vasomodulator and immunomodulators. Imunomudulator could act as immunosuppressive factors that can suppress nonspecific immune system of the host and modulate the change of T helper 1 (Th1) toward T helper 2 (Th2) response, which is advantageous for malaria parasite to infect human host. This research wanted to evaluate human immune respons in endemic area against salivary gland protein extract (SGPE) from its major malaria vector i.e. Anopheles sundaicus (An. sundaicus). Analysis of human immune response was conducted quantitatively by ELISA (Enzyme Link Immunosorbend Assay) towards IgG from human sera samples after cross reacted with SGPE. The results showed that exposures to An. sundaicus were able to induce high levels of IgG. IgG anti salivary proteins of An. sundaicus is higher than the levels of IgG anti salivary proteins of Ae. aegypti. Furthermore, the age group 11-40 years with the highest bites probability, had the highest IgG levels compared to other age groups.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"34 1","pages":"4-4"},"PeriodicalIF":0.0,"publicationDate":"2017-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78304916","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}
Tempeh is a traditional fermented soybean product from Indonesia. Although tempeh is consumed as daily menu in Indonesia, its nutrigenomic study employing human has not been reported yet. On the other hand, our study in mice showed that tempeh could enhance immune system, especially by increasing secretory immunoglobulin A production in ileum and colon. Tempeh was also found to be potential in modulating the composition of gut microbiota. Therefore, the objective of this study was to analyze the impact of tempeh supplementation on the profiles of human intestinal immune system and gut microbiota analysis. This experimental design was reviewed and approved by the ethics committee. A total of 16 participants, comprising of each 8 healthy females and males, aged between 20 and 23 were recruited to this study. The volunteers consumed 200 mL milk from day 1-8 followed by consumption of 100 grams steamed tempeh each day from day 9-24. Fecal samples, which were taken on day 9 and 25, were analyzed with half sandwich ELISA for IgA enumeration while fecal samples, which were taken on day 0, 9, and 25, were analyzed for Akkermansia muciniphila enumeration employing quantitative real time PCR. The result of this study suggesting that tempeh supplementation might act as paraprobiotic and slimming agent since tempeh enhanced production of IgA and increased the number of A. muciniphila in human intestinal tract.
{"title":"Effect of Tempeh Supplementation on the Profiles of Human Intestinal Immune System and Gut Microbiota","authors":"S. Stéphanie, N. K. Ratih, Susan Soka, A. Suwanto","doi":"10.5454/MI.11.1.2","DOIUrl":"https://doi.org/10.5454/MI.11.1.2","url":null,"abstract":"Tempeh is a traditional fermented soybean product from Indonesia. Although tempeh is consumed as daily menu in Indonesia, its nutrigenomic study employing human has not been reported yet. On the other hand, our study in mice showed that tempeh could enhance immune system, especially by increasing secretory immunoglobulin A production in ileum and colon. Tempeh was also found to be potential in modulating the composition of gut microbiota. Therefore, the objective of this study was to analyze the impact of tempeh supplementation on the profiles of human intestinal immune system and gut microbiota analysis. This experimental design was reviewed and approved by the ethics committee. A total of 16 participants, comprising of each 8 healthy females and males, aged between 20 and 23 were recruited to this study. The volunteers consumed 200 mL milk from day 1-8 followed by consumption of 100 grams steamed tempeh each day from day 9-24. Fecal samples, which were taken on day 9 and 25, were analyzed with half sandwich ELISA for IgA enumeration while fecal samples, which were taken on day 0, 9, and 25, were analyzed for Akkermansia muciniphila enumeration employing quantitative real time PCR. The result of this study suggesting that tempeh supplementation might act as paraprobiotic and slimming agent since tempeh enhanced production of IgA and increased the number of A. muciniphila in human intestinal tract.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"22 1","pages":"2-2"},"PeriodicalIF":0.0,"publicationDate":"2017-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88610072","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}
Nitric acid mutations are known could be used for strain improvement. This research aimed to studies IAA production by nitric acid mutan were compared with wild type. Mutation were conducted with some different treatment time such as 0, 30, 60, 90 and 120 min subsequently it were measured for IAA production. Isolate H6 as -1wild type isolates were also molecularly identified. The wild strain exhibited 53.83 µg mL of IAA while the -1 -1. nitric acid mutan within a range 77.39 µg mL to 95.70 µg mL Isolates H6.60 exhibited the highest IAA -1 production which 39.87 µg mL higher were compared with wild-type. Based on 16S rRNA gene analysis, isolate H6 belonged to Lysobacter sp. ES2-22.
{"title":"Studies for IAA (Indole-3-Acetic Acid) Production by Isolates H6 with Nitric Acid Mutation","authors":"R. F. W. Putrie, T. Widowati, H. Sukiman","doi":"10.5454/MI.11.1.3","DOIUrl":"https://doi.org/10.5454/MI.11.1.3","url":null,"abstract":"Nitric acid mutations are known could be used for strain improvement. This research aimed to studies IAA production by nitric acid mutan were compared with wild type. Mutation were conducted with some different treatment time such as 0, 30, 60, 90 and 120 min subsequently it were measured for IAA production. Isolate H6 as -1wild type isolates were also molecularly identified. The wild strain exhibited 53.83 µg mL of IAA while the -1 -1. nitric acid mutan within a range 77.39 µg mL to 95.70 µg mL Isolates H6.60 exhibited the highest IAA -1 production which 39.87 µg mL higher were compared with wild-type. Based on 16S rRNA gene analysis, isolate H6 belonged to Lysobacter sp. ES2-22.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"12 1","pages":"3-3"},"PeriodicalIF":0.0,"publicationDate":"2017-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79149615","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}
M. Cahyani, I. Helianti, N. Nurhayati, A. Abinawanto
Lipases (EC 3.1.1.3) are classified as hydrolases that hydrolyze lipids. These enzymes have potential application in biotechnology and industrial process. In previous study we have cloned the synthetic Rhizomucor miehei lipase gene using the vector pUC57 in Escherichia coli DH5α, but only found the very low enzymes activity. This study aimed to clone Rhizomucor miehei synthetic lipase gene into Pichia pastoris expression plasmid for lipase expression with the original signal peptide. A DNA fragment with the original signal peptide had been obtained by PCR, cut by Xho I and Xba I and then ligated into pPICZα A linearized with the same enzymes. The mixture of ligation, then was transformed into Escherichia coli DH5α. Zeocin-resistant transformants were selected and contained plasmid was analyzed by restriction enzymes analyses, PCR, and DNA sequenced. As the result, a Rhizomucor miehei lipase gene (RMlip) with the size of 1132 bp was successfully cloned to pPICZα A. The recombinant plasmid with the correct DNA sequence was transformed into Pichia pastoris X33. Cultivation of recombinant P. pastoris was carried out with the addition of 1.5% methanol every day with appropriate aeration. The recombinant lipase produced by Pichia pastoris X33 containing RMlip oin its chromosomal DNA had optimal temperature and pH 30 C and 9.0, respectively.
{"title":"Cloning of Synthetic Lipase Gene from Rhizomucor miehei with Original Signal Peptide in Pichia pastoris","authors":"M. Cahyani, I. Helianti, N. Nurhayati, A. Abinawanto","doi":"10.5454/MI.11.1.1","DOIUrl":"https://doi.org/10.5454/MI.11.1.1","url":null,"abstract":"Lipases (EC 3.1.1.3) are classified as hydrolases that hydrolyze lipids. These enzymes have potential application in biotechnology and industrial process. In previous study we have cloned the synthetic Rhizomucor miehei lipase gene using the vector pUC57 in Escherichia coli DH5α, but only found the very low enzymes activity. This study aimed to clone Rhizomucor miehei synthetic lipase gene into Pichia pastoris expression plasmid for lipase expression with the original signal peptide. A DNA fragment with the original signal peptide had been obtained by PCR, cut by Xho I and Xba I and then ligated into pPICZα A linearized with the same enzymes. The mixture of ligation, then was transformed into Escherichia coli DH5α. Zeocin-resistant transformants were selected and contained plasmid was analyzed by restriction enzymes analyses, PCR, and DNA sequenced. As the result, a Rhizomucor miehei lipase gene (RMlip) with the size of 1132 bp was successfully cloned to pPICZα A. The recombinant plasmid with the correct DNA sequence was transformed into Pichia pastoris X33. Cultivation of recombinant P. pastoris was carried out with the addition of 1.5% methanol every day with appropriate aeration. The recombinant lipase produced by Pichia pastoris X33 containing RMlip oin its chromosomal DNA had optimal temperature and pH 30 C and 9.0, respectively.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"1 1","pages":"1-10"},"PeriodicalIF":0.0,"publicationDate":"2017-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90082167","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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