Pub Date : 2023-02-10DOI: 10.35118/apjmbb.2023.031.1.02
'Aina Nabilah Faizah Ahmad Bustamam, Nur Sazwani Daud, Zaheda Mohamad Azam, M. A. Rosli, Solleh Ramli, N. Zainol, M. Nadri, H. Leong, N. Othman
Exopolysaccharides (EPS) are well-known biopolymers secreted by several lactic acid bacteria with combination of various strains. The aim of this study is to increase EPS production by co-culturing Lactobacillus plantarum ATCC 8014 with Saccharomyces cerevisiae ICA-Y01 and study the changes in the functional characteristics of the EPS from both cultivations. In this study, the production and functional characteristics of EPS from co-cultivation culture of L. plantarum ATCC 8014 with S. cerevisiae ICA-Y01 were evaluated. The co-cultivation of L. plantarum ATCC 8014 with S. cerevisiae ICA-Y01was markedly increased EPS production up to 55.84% with 6.8 g/l yield after 20 hours cultivation. The pH of the co-cultivation culture was remained constantly at 5.2 until the end of cultivation. Furthermore, co-cultivation under pH 6 in the 16L bioreactor showed a higher growth rate of 0.214 h-1 and EPS production increased up to 104.44% when compared with single cultivation of L. plantarum ATCC 8014. This result clearly indicates the importance of growing the cells in the controlled pH condition when cultivated with S. cerevisiae ICA-Y01 to enhance EPS production. The functional characteristics of EPS secreted from both cultivation strategies were also evaluated. FT-IR spectroscopy confirmed EPS presence from both cultivations, indicating functional group of the polysaccharide with D-glucose units bound by α-(1→6). The EPS production from single cultivation showed a higher DPPH radical scavenging activity (88.21%) and IC50 (19.57%) as compared to EPS produced from co-cultivation with DPPH scavenging exhibited 32.45% with no IC50 value detected. Furthermore, solubility and water uptake of EPS from single cultivation are higher in comparison to co-cultivation. In conclusion, higher efficiency in the bioactivity of EPS from the single cultivation of L. plantarum ATCC 8014 was confirmed even though the EPS yield is low as compared to EPS synthesis through inter-kingdom cultivation.
胞外多糖(Exopolysaccharides, EPS)是由多种乳酸菌结合多种菌株分泌的生物聚合物。本研究旨在通过植物乳杆菌ATCC 8014与酿酒酵母ICA-Y01共培养提高EPS产量，并研究两种培养下EPS功能特性的变化。研究了L. plantarum ATCC 8014与S. cerevisiae ICA-Y01共培养EPS的产量和功能特性。l . plantarum ATCC 8014与S. cerevisiae ica - y01共培养20 h后EPS产量显著提高，达55.84%，产量为6.8 g/l。共培养的pH值一直保持在5.2，直到培养结束。此外，在pH为6的16L生物反应器中，与单独培养相比，L. plantarum ATCC 8014的生长速率为0.214 h-1, EPS产量提高了104.44%。这一结果清楚地说明了酵母ICA-Y01在控制pH条件下培养细胞对提高EPS产量的重要性。并对两种培养方式分泌的EPS的功能特性进行了评价。FT-IR光谱证实了两种培养物均存在EPS，表明多糖的官能团与α-(1→6)结合的d -葡萄糖单元。单独培养产生的EPS清除DPPH自由基的活性为88.21%，IC50为19.57%，而联合培养产生的EPS清除DPPH自由基的活性为32.45%，IC50未检测到。此外，单独培养EPS的溶解度和吸水率高于共培养。综上所述，与跨界栽培合成EPS相比，单株栽培的EPS产量较低，但单株栽培的EPS生物活性更高。
{"title":"Production and functional characteristics of exopolysaccharide by Lactobacillus plantarum co-cultivation with Saccharomyces cerevisiae","authors":"'Aina Nabilah Faizah Ahmad Bustamam, Nur Sazwani Daud, Zaheda Mohamad Azam, M. A. Rosli, Solleh Ramli, N. Zainol, M. Nadri, H. Leong, N. Othman","doi":"10.35118/apjmbb.2023.031.1.02","DOIUrl":"https://doi.org/10.35118/apjmbb.2023.031.1.02","url":null,"abstract":"Exopolysaccharides (EPS) are well-known biopolymers secreted by several lactic acid bacteria with combination of various strains. The aim of this study is to increase EPS production by co-culturing Lactobacillus plantarum ATCC 8014 with Saccharomyces cerevisiae ICA-Y01 and study the changes in the functional characteristics of the EPS from both cultivations. In this study, the production and functional characteristics of EPS from co-cultivation culture of L. plantarum ATCC 8014 with S. cerevisiae ICA-Y01 were evaluated. The co-cultivation of L. plantarum ATCC 8014 with S. cerevisiae ICA-Y01was markedly increased EPS production up to 55.84% with 6.8 g/l yield after 20 hours cultivation. The pH of the co-cultivation culture was remained constantly at 5.2 until the end of cultivation. Furthermore, co-cultivation under pH 6 in the 16L bioreactor showed a higher growth rate of 0.214 h-1 and EPS production increased up to 104.44% when compared with single cultivation of L. plantarum ATCC 8014. This result clearly indicates the importance of growing the cells in the controlled pH condition when cultivated with S. cerevisiae ICA-Y01 to enhance EPS production. The functional characteristics of EPS secreted from both cultivation strategies were also evaluated. FT-IR spectroscopy confirmed EPS presence from both cultivations, indicating functional group of the polysaccharide with D-glucose units bound by α-(1→6). The EPS production from single cultivation showed a higher DPPH radical scavenging activity (88.21%) and IC50 (19.57%) as compared to EPS produced from co-cultivation with DPPH scavenging exhibited 32.45% with no IC50 value detected. Furthermore, solubility and water uptake of EPS from single cultivation are higher in comparison to co-cultivation. In conclusion, higher efficiency in the bioactivity of EPS from the single cultivation of L. plantarum ATCC 8014 was confirmed even though the EPS yield is low as compared to EPS synthesis through inter-kingdom cultivation.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76937312","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}
Pub Date : 2023-01-19DOI: 10.35118/apjmbb.2023.031.1.01
Nur Fathiah Rosilan, M. Samsulrizal, Norsyahida Rani, N. Samsulrizal, Z. Zainuddin, T. Sundram
Stevia rebaudiana is a plant under the Asteraceae family and has been reported as a healthier alternative to sugar. Steviol glycosides (SGs) is the group of secondary metabolites responsible for the sweet taste. Among nine SGs synthesised by S. rebaudiana, stevioside and rebaudioside A are the sweetest. The biosynthetic pathway of SGs partly involves conversion of geranylgeranyl diphosphate (GGDP) into steviol, catalysed by ent- kaurene synthase (KS), ent-copalyl diphosphate synthase (CPPS), and kaurene oxidase (KO). This study focuses on in silico molecular characterization and phylogenetic analysis of KS from Malaysia’s S. rebaudiana MS007 variety (Stevia MS007). The transcriptomic dataset of S. rebaudiana accession MS007 was used in initial experiment toward analysing the KS. Through the blastx homology search using transcriptomic dataset query Cluster-31069.42907, the Stevia rebaudiana kaurene synthase (SrKS) sequence was identified with the highest similarity percentage identity (99.62%). The protein domain prediction using InterPro yields IPR005630 (terpene synthase metal-binding domain) at positions 490 to 755 and IPR001906 (terpene synthase-N-terminal-domain) at positions 258 to 477. Multiple sequence alignment was conducted using MUSCLE and MEGA-X as phylogenetic tree analysis tool for constructing the phylogenetic analysis tree. Based on the bootstrap value from the phylogenetic analysis, Cluster-31069.42907 represents relationships between the ancestors. Since both Helianthus annuus and S. rebaudiana are Asteraceae species, the bootstrap value for both species was 100%. In conclusion, this research contributes to a better understanding of Stevia MS007 KS via in silico analysis.
{"title":"Molecular characterization and phylogenetic analysis of kaurene synthase protein in Stevia rebaudiana MS007","authors":"Nur Fathiah Rosilan, M. Samsulrizal, Norsyahida Rani, N. Samsulrizal, Z. Zainuddin, T. Sundram","doi":"10.35118/apjmbb.2023.031.1.01","DOIUrl":"https://doi.org/10.35118/apjmbb.2023.031.1.01","url":null,"abstract":"Stevia rebaudiana is a plant under the Asteraceae family and has been reported as a healthier alternative to sugar. Steviol glycosides (SGs) is the group of secondary metabolites responsible for the sweet taste. Among nine SGs synthesised by S. rebaudiana, stevioside and rebaudioside A are the sweetest. The biosynthetic pathway of SGs partly involves conversion of geranylgeranyl diphosphate (GGDP) into steviol, catalysed by ent- kaurene synthase (KS), ent-copalyl diphosphate synthase (CPPS), and kaurene oxidase (KO). This study focuses on in silico molecular characterization and phylogenetic analysis of KS from Malaysia’s S. rebaudiana MS007 variety (Stevia MS007). The transcriptomic dataset of S. rebaudiana accession MS007 was used in initial experiment toward analysing the KS. Through the blastx homology search using transcriptomic dataset query Cluster-31069.42907, the Stevia rebaudiana kaurene synthase (SrKS) sequence was identified with the highest similarity percentage identity (99.62%). The protein domain prediction using InterPro yields IPR005630 (terpene synthase metal-binding domain) at positions 490 to 755 and IPR001906 (terpene synthase-N-terminal-domain) at positions 258 to 477. Multiple sequence alignment was conducted using MUSCLE and MEGA-X as phylogenetic tree analysis tool for constructing the phylogenetic analysis tree. Based on the bootstrap value from the phylogenetic analysis, Cluster-31069.42907 represents relationships between the ancestors. Since both Helianthus annuus and S. rebaudiana are Asteraceae species, the bootstrap value for both species was 100%. In conclusion, this research contributes to a better understanding of Stevia MS007 KS via in silico analysis.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80022696","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}
Pub Date : 2023-01-03DOI: 10.35118/apjmbb.2022.030.4.08
J. Arulraj, Ashok Raj Kattur Venkatachalam, Revathy Soundararajan, Rajesh Mani
Flocculation is used to effectively separate suspended colloids in domestic and industrial wastewater. Flocculants are classified into three categories as organic, inorganic and natural flocculants. Its flocculating properties, ease of use and commercial use have led to the widespread use of organic and inorganic flocculants. However, it can cause serious health problems due to its carcinogenicity and neurotoxicity. Therefore, natural bioflocculants are used to treat wastewater without harming humans and the environment. Natural flocculants are non-toxic, environmentally friendly and capable of flotation even at low concentrations. This article also discusses the classification, functions, mechanisms and applications of flocculants. Applications of natural flocculants and flocculation efficiency in the treatment of industrial wastes such as food, heavy metal and dyeing are discussed. Future studies will use methods to understand how agricultural and food wastes are used for cost-effective bioflocculant production. Bacterial consortia and new novel marine bacteria are indicated for large-scale industrial production.
{"title":"Microbial flocculants as an excellent alternative to synthetic flocculants for industrial application: A comprehensive review","authors":"J. Arulraj, Ashok Raj Kattur Venkatachalam, Revathy Soundararajan, Rajesh Mani","doi":"10.35118/apjmbb.2022.030.4.08","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.08","url":null,"abstract":"Flocculation is used to effectively separate suspended colloids in domestic and industrial wastewater. Flocculants are classified into three categories as organic, inorganic and natural flocculants. Its flocculating properties, ease of use and commercial use have led to the widespread use of organic and inorganic flocculants. However, it can cause serious health problems due to its carcinogenicity and neurotoxicity. Therefore, natural bioflocculants are used to treat wastewater without harming humans and the environment. Natural flocculants are non-toxic, environmentally friendly and capable of flotation even at low concentrations. This article also discusses the classification, functions, mechanisms and applications of flocculants. Applications of natural flocculants and flocculation efficiency in the treatment of industrial wastes such as food, heavy metal and dyeing are discussed. Future studies will use methods to understand how agricultural and food wastes are used for cost-effective bioflocculant production. Bacterial consortia and new novel marine bacteria are indicated for large-scale industrial production.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74640211","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}
Pub Date : 2022-12-28DOI: 10.35118/apjmbb.2022.030.4.07
Manjunatha Bukkambudi Krishnaswamy, V. Kanagasabapathy, A. Gomathi, Dr. Ramachandrappa, Pratheeksha Gurumurthy, R. Kumar, Urvi Narayan, Krithika Shanmugam, Girinath G. Pillai
Immune cell malignancy such as Acute T- cell Lymphoblastic Leukaemia is generally associated with high rate of relapse and often does not respond to salvage therapy. Thus, identification of novel treatment regimens or cell apoptosis pathways and therapeutic agents without major side effects is necessary. TRAIL-induced apoptotic pathway is one such pathway that is usually blocked by anti-apoptotic proteins like Bcl-2. This research estimated and compared the ability of few common flavonoids to re-activate TRAIL-induced apoptosis by blocking Bcl-2 protein. Studies were carried out to understand the interaction between binding energy of the Flavonoids with Bcl-2 protein in cancer cells. The pharmacokinetic and toxicity profiling was performed to study the potency of the flavonoids as a lead candidate. Baicalein was selected as lead molecule because of its lower binding energy and its ability to increase Mitochondrial Membrane Potential as studied from its ADME properties. For validation of apoptosis of Baicalein by TRAIL-induced owing to Bcl-2 analysis of cell cycle and Gene expression studies were carried out on Jurkat T cells.
{"title":"Interaction studies of flavonoids with Bcl-2 protein to re-activate apoptosis in JurkatT-cells by induced TRAIL","authors":"Manjunatha Bukkambudi Krishnaswamy, V. Kanagasabapathy, A. Gomathi, Dr. Ramachandrappa, Pratheeksha Gurumurthy, R. Kumar, Urvi Narayan, Krithika Shanmugam, Girinath G. Pillai","doi":"10.35118/apjmbb.2022.030.4.07","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.07","url":null,"abstract":"Immune cell malignancy such as Acute T- cell Lymphoblastic Leukaemia is generally associated with high rate of relapse and often does not respond to salvage therapy. Thus, identification of novel treatment regimens or cell apoptosis pathways and therapeutic agents without major side effects is necessary. TRAIL-induced apoptotic pathway is one such pathway that is usually blocked by anti-apoptotic proteins like Bcl-2. This research estimated and compared the ability of few common flavonoids to re-activate TRAIL-induced apoptosis by blocking Bcl-2 protein. Studies were carried out to understand the interaction between binding energy of the Flavonoids with Bcl-2 protein in cancer cells. The pharmacokinetic and toxicity profiling was performed to study the potency of the flavonoids as a lead candidate. Baicalein was selected as lead molecule because of its lower binding energy and its ability to increase Mitochondrial Membrane Potential as studied from its ADME properties. For validation of apoptosis of Baicalein by TRAIL-induced owing to Bcl-2 analysis of cell cycle and Gene expression studies were carried out on Jurkat T cells.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76546469","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}
Pub Date : 2022-12-24DOI: 10.35118/apjmbb.2022.030.4.03
Siti Fatimah Suboh, A. M. Mahat, M. H. Yusof, Wan Razarinah Wan Abdul Razak
Strobilanthes crispus is a plant species from the family of Acanthaceae, known as ‘pecah kaca’ or ‘jin batu’ among Malaysians. S. crispus have medicinal properties that are anti-diabetic, diuretic, laxative, antimicrobial and wound healing properties. The study aimed to investigate the antimicrobial activity of S. crispus leaves aqueous extract and iron oxide nanoparticles (IONP) against the selected human pathogens namely Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Bacillus cereus ATCC 11778, Salmonella typhi ATCC 14028, Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404. The antimicrobial properties of S. crispus leaves aqueous extract and IONP was measured through the disc diffusion test. The inhibitory action of S. crispus leaves aqueous extract was only visible against E. coli; at 25 mg/ml with mean diameter zone of inhibition was 7.3 ± 0.017 mm and at 50 mg/ml (8.7 ± 0.017 mm). However, IONP was able to show better antimicrobial activity against all selected human pathogens compared to S. crispus leaves aqueous extract. The largest mean diameter zone of inhibition exhibited was 11.50 ± 0.288 mm at 0.2 M IONP and 9.00 ± 0.577 mm at 0.3 M IONP against S. aureus. The properties and structural of IONP was investigated through UV-Vis, SEM, XRD and FTIR method. Hence, these data concluded that S. crispus leaves aqueous extract and IONP have antimicrobial properties against selected human pathogens thus act as potential antimicrobial agent in pharmaceutical industry.
{"title":"Antimicrobial activity of Strobilanthes crispus leaves aqueous extract and green biosynthesis iron oxide nanoparticles against selected human pathogens","authors":"Siti Fatimah Suboh, A. M. Mahat, M. H. Yusof, Wan Razarinah Wan Abdul Razak","doi":"10.35118/apjmbb.2022.030.4.03","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.03","url":null,"abstract":"Strobilanthes crispus is a plant species from the family of Acanthaceae, known as ‘pecah kaca’ or ‘jin batu’ among Malaysians. S. crispus have medicinal properties that are anti-diabetic, diuretic, laxative, antimicrobial and wound healing properties. The study aimed to investigate the antimicrobial activity of S. crispus leaves aqueous extract and iron oxide nanoparticles (IONP) against the selected human pathogens namely Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Bacillus cereus ATCC 11778, Salmonella typhi ATCC 14028, Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404. The antimicrobial properties of S. crispus leaves aqueous extract and IONP was measured through the disc diffusion test. The inhibitory action of S. crispus leaves aqueous extract was only visible against E. coli; at 25 mg/ml with mean diameter zone of inhibition was 7.3 ± 0.017 mm and at 50 mg/ml (8.7 ± 0.017 mm). However, IONP was able to show better antimicrobial activity against all selected human pathogens compared to S. crispus leaves aqueous extract. The largest mean diameter zone of inhibition exhibited was 11.50 ± 0.288 mm at 0.2 M IONP and 9.00 ± 0.577 mm at 0.3 M IONP against S. aureus. The properties and structural of IONP was investigated through UV-Vis, SEM, XRD and FTIR method. Hence, these data concluded that S. crispus leaves aqueous extract and IONP have antimicrobial properties against selected human pathogens thus act as potential antimicrobial agent in pharmaceutical industry.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84693594","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}
Pub Date : 2022-12-01DOI: 10.35118/apjmbb.2022.030.4.05
M. Azmi, Alyaa Al Khateeb, S. Ab Rahim, G. Froemming, E. Omar
Nisin, a known food preservative, has antimicrobial and anticancer properties. Nisin is a polycyclic polypeptide with five internal rings; a feature that strongly influence the choice of solvent to attain effectivity of the compound for all cell culture experiments. Previous studies used a number of solvents but comparison between these to identify the best solvent have not been reported. Aim: To investigate the effect of nisin ZP (NZP) when dissolved in four different solvents on the viability of MG63 osteosarcoma cells. Materials and methods: MG-63 cells were incubated overnight at 37°C, 5% CO2 and treated with increasing concentration of dimethyl sulfoxide (DMSO), methanol and acetic acid to observe the cytotoxic effects of these chemicals onto the cells via MTS assay. NZP was then dissolved in distilled water and the three solvents in increasing concentrations and applied to the cultured cells. Cell viability and morphology were documented. Results: Acetic acid, DMSO and methanol at 0.05% maintained more than 90% viability of MG-63. NZP precipitated at higher concentrations (≥ 800µg/ml) when dissolved in DMSO, methanol and distilled water which interfered with the measurement of formazan density in the cell viability assay. Increasing doses of NZP in 0.05% acetic acid showed the least interference with the desirable reduction of the cancer cell viability. Microscopically, as concentration of NZP increases, cell shrinkage and apoptotic bodies were more visible. In addition, brown precipitates were abundantly observed. Conclusion: Acetic acid provides the optimal solubility of NZP and the best solvent to use in experiments involving MG-63 cells.
{"title":"Effect of different solvents on nisin ZP potential as anticancer agent against MG-63 osteosarcoma cells","authors":"M. Azmi, Alyaa Al Khateeb, S. Ab Rahim, G. Froemming, E. Omar","doi":"10.35118/apjmbb.2022.030.4.05","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.05","url":null,"abstract":"Nisin, a known food preservative, has antimicrobial and anticancer properties. Nisin is a polycyclic polypeptide with five internal rings; a feature that strongly influence the choice of solvent to attain effectivity of the compound for all cell culture experiments. Previous studies used a number of solvents but comparison between these to identify the best solvent have not been reported. Aim: To investigate the effect of nisin ZP (NZP) when dissolved in four different solvents on the viability of MG63 osteosarcoma cells. Materials and methods: MG-63 cells were incubated overnight at 37°C, 5% CO2 and treated with increasing concentration of dimethyl sulfoxide (DMSO), methanol and acetic acid to observe the cytotoxic effects of these chemicals onto the cells via MTS assay. NZP was then dissolved in distilled water and the three solvents in increasing concentrations and applied to the cultured cells. Cell viability and morphology were documented. Results: Acetic acid, DMSO and methanol at 0.05% maintained more than 90% viability of MG-63. NZP precipitated at higher concentrations (≥ 800µg/ml) when dissolved in DMSO, methanol and distilled water which interfered with the measurement of formazan density in the cell viability assay. Increasing doses of NZP in 0.05% acetic acid showed the least interference with the desirable reduction of the cancer cell viability. Microscopically, as concentration of NZP increases, cell shrinkage and apoptotic bodies were more visible. In addition, brown precipitates were abundantly observed. Conclusion: Acetic acid provides the optimal solubility of NZP and the best solvent to use in experiments involving MG-63 cells.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75536617","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}
Pub Date : 2022-12-01DOI: 10.35118/apjmbb.2022.030.4.06
P. Lestari, T. P. Priyatno, K. Nugroho, R. T. Terryana, Mastur, I. Manzila, Andiningtyas Mula Pertiwi, Agus Tri Sudaryanto, B. Tiesnamurti
Multiple births or twinning in cattle is viewed as a natural phenomenon that still needs to be explored in the genetic aspect. The occurrence of twinning birth in cattle has been found in Indonesia both on beef and dairy breeds, however, the frequency is very low. This study aimed to identify the genetic variation of the candidate loci of Bovine chromosome 10 (BTA10) to control twinning in beef and dairy cattle in Indonesia. At least 57 historical twin births of both beef and dairy breeds across Central Java with the highest twins in cattle, were identified in 2017 during the dry season. The ratio of twin calves was relatively similar between female and male calves. Male twins showed the highest number (37.8%), while freemartin heifers were also highly born (28.9%) among twin births. Forty-nine heads out of 152 heads were sequenced on the partial regions in BTA10 that were highly homologous with the ovulation rate gene. DNA sequences polymorphism was identified at three regions in the BTA10 among twins of Simmental, Ongole, Limousin, Simmental-Ongole, and Holstein Friesian cattle. A significant association was not revealed between identified SNP and twinning birth potential in all breeds. Notably, one SNP (A/G) at 410 nucleotides in the consensus sequence of BTA10-2 appeared to be highly significantly associated with twinning birth potential in beef cattle of the Simmental. Taken together, this SNP would be a potential genetic marker for early selection of twinning birth which is promising for dramatic improvement of beef cattle production.
{"title":"Genetic variation of the candidate loci controlling twinning in the beef and dairy cattle breeds in Indonesia","authors":"P. Lestari, T. P. Priyatno, K. Nugroho, R. T. Terryana, Mastur, I. Manzila, Andiningtyas Mula Pertiwi, Agus Tri Sudaryanto, B. Tiesnamurti","doi":"10.35118/apjmbb.2022.030.4.06","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.06","url":null,"abstract":"Multiple births or twinning in cattle is viewed as a natural phenomenon that still needs to be explored in the genetic aspect. The occurrence of twinning birth in cattle has been found in Indonesia both on beef and dairy breeds, however, the frequency is very low. This study aimed to identify the genetic variation of the candidate loci of Bovine chromosome 10 (BTA10) to control twinning in beef and dairy cattle in Indonesia. At least 57 historical twin births of both beef and dairy breeds across Central Java with the highest twins in cattle, were identified in 2017 during the dry season. The ratio of twin calves was relatively similar between female and male calves. Male twins showed the highest number (37.8%), while freemartin heifers were also highly born (28.9%) among twin births. Forty-nine heads out of 152 heads were sequenced on the partial regions in BTA10 that were highly homologous with the ovulation rate gene. DNA sequences polymorphism was identified at three regions in the BTA10 among twins of Simmental, Ongole, Limousin, Simmental-Ongole, and Holstein Friesian cattle. A significant association was not revealed between identified SNP and twinning birth potential in all breeds. Notably, one SNP (A/G) at 410 nucleotides in the consensus sequence of BTA10-2 appeared to be highly significantly associated with twinning birth potential in beef cattle of the Simmental. Taken together, this SNP would be a potential genetic marker for early selection of twinning birth which is promising for dramatic improvement of beef cattle production.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79803760","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}
Pub Date : 2022-11-24DOI: 10.35118/apjmbb.2022.030.4.04
N. Hoang, Nguyen Tuan Anh, T. Hieu, L. T. Ngan
Clarithromycin (CLA) and levofloxacin (LFX) have been recommended as the most effective antibiotics for treating Helicobacter pylori infection. However, the increase in H. pylori’s resistance to antibiotics is an alarming and growing challenge. The study aimed to determine the antimicrobial resistance profiles of H. pylori strains isolated from patients at the Hospital for Tropical Diseases and to detect point mutations in 23S ribosomal RNA (23S rRNA) and gyraseA (gyrA) genes. Point mutations in 23S rRNA and gyrA were detected using Sanger sequencing. Antibiotic resistance was tested by the microdilution method. Out of the 45 isolates, 44 (97.8%) were found to be resistant to at least one antibiotic, 38 (84.4%) resistant to metronidazole (MTZ), and all were sensitive to amoxicillin (AMX) and tetracycline (TET). Sixty percent of the isolated strains were resistant to 3-6 antibiotics; among them, multiple drug resistance (MDR) strains presented twenty percent resistance to more than two classes of antibiotics. Eleven strains (24.4%) carried two mutations associated with CLA and LFX but only nine of them were dual resistant to CLA and LFX, and twenty-four (53.3%) carried either CLA or LFX mutation. The point mutations A2143G in the 23S rRNA and N87K (Asn87Lys) in the gyrA were detected in the CLA and LFX resistant strains, respectively. The prevalence of MDR, especially CLA, MTZ, and LFX resistance, in the H. pylori isolates suggests that the use of these antibiotics need to be more considerable and cautious.
{"title":"Evaluation of clarithromycin and levofloxacin resistance of Helicobacter pylori strains isolated from patients at the Hospital for Tropical Diseases, Ho Chi Minh City","authors":"N. Hoang, Nguyen Tuan Anh, T. Hieu, L. T. Ngan","doi":"10.35118/apjmbb.2022.030.4.04","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.04","url":null,"abstract":"Clarithromycin (CLA) and levofloxacin (LFX) have been recommended as the most effective antibiotics for treating Helicobacter pylori infection. However, the increase in H. pylori’s resistance to antibiotics is an alarming and growing challenge. The study aimed to determine the antimicrobial resistance profiles of H. pylori strains isolated from patients at the Hospital for Tropical Diseases and to detect point mutations in 23S ribosomal RNA (23S rRNA) and gyraseA (gyrA) genes. Point mutations in 23S rRNA and gyrA were detected using Sanger sequencing. Antibiotic resistance was tested by the microdilution method. Out of the 45 isolates, 44 (97.8%) were found to be resistant to at least one antibiotic, 38 (84.4%) resistant to metronidazole (MTZ), and all were sensitive to amoxicillin (AMX) and tetracycline (TET). Sixty percent of the isolated strains were resistant to 3-6 antibiotics; among them, multiple drug resistance (MDR) strains presented twenty percent resistance to more than two classes of antibiotics. Eleven strains (24.4%) carried two mutations associated with CLA and LFX but only nine of them were dual resistant to CLA and LFX, and twenty-four (53.3%) carried either CLA or LFX mutation. The point mutations A2143G in the 23S rRNA and N87K (Asn87Lys) in the gyrA were detected in the CLA and LFX resistant strains, respectively. The prevalence of MDR, especially CLA, MTZ, and LFX resistance, in the H. pylori isolates suggests that the use of these antibiotics need to be more considerable and cautious.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82258904","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}
Pub Date : 2022-11-15DOI: 10.35118/apjmbb.2022.030.4.02
M. Gondokesumo, L. Sapei, M. Wahjudi, N. Suseno, T. Adiarto
Virgin coconut oil (VCO) is a natural product that contains mostly medium-chain lipids, such as palmitates, stearates, and oleates. This study aims to explore whether VCO would make an effective to Nsp3b as one of target for virtual ligand screening of potential drug targets inhibitor of SARS-CoV-2, especially medium-chain content. In this study, computational investigations (in silico) were conducted using five long-chain molecules constituting VCO, namely palmitate, myristate, stearate, laurate, and oleate. Molecular docking simulation was conducted using the PLANTS 1.1. The binding affinity results revealed palmitate, and stearate have lower score than the co-crystalize ligand of Nsp3. Stearate and palmitate binding affinity score were -6.45 and -6.23 respectively, while co-crystalize ligand as our ligand control is -5.71, despite co-crystalize ligand hydrogen bonds is more than both of palmitate and stearate. In addition to molecular docking, we perform molecular dynamic simulation and found stearate relatively stable to bind Nsp3. The RMSD of complex protein to stearate was stable below 1 nm over 20 ns simulation. This could be caused by hydrogen bonds between stearate and Nsp3 protein, where average of hydrogen bond is 1.2, and recorded to be higher during the last 10 ns with an average of 1.5. Both palmitate and stearate also found have biological activity against several virus including adenovirus, poxvirus, and influenza virus with score greater than 0.5 (score from 0 to 1).
{"title":"Molecular mechanism of virgin coconut oil as a Nsp-3 inhibitor of SARS-CoV-2","authors":"M. Gondokesumo, L. Sapei, M. Wahjudi, N. Suseno, T. Adiarto","doi":"10.35118/apjmbb.2022.030.4.02","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.02","url":null,"abstract":"Virgin coconut oil (VCO) is a natural product that contains mostly medium-chain lipids, such as palmitates, stearates, and oleates. This study aims to explore whether VCO would make an effective to Nsp3b as one of target for virtual ligand screening of potential drug targets inhibitor of SARS-CoV-2, especially medium-chain content. In this study, computational investigations (in silico) were conducted using five long-chain molecules constituting VCO, namely palmitate, myristate, stearate, laurate, and oleate. Molecular docking simulation was conducted using the PLANTS 1.1. The binding affinity results revealed palmitate, and stearate have lower score than the co-crystalize ligand of Nsp3. Stearate and palmitate binding affinity score were -6.45 and -6.23 respectively, while co-crystalize ligand as our ligand control is -5.71, despite co-crystalize ligand hydrogen bonds is more than both of palmitate and stearate. In addition to molecular docking, we perform molecular dynamic simulation and found stearate relatively stable to bind Nsp3. The RMSD of complex protein to stearate was stable below 1 nm over 20 ns simulation. This could be caused by hydrogen bonds between stearate and Nsp3 protein, where average of hydrogen bond is 1.2, and recorded to be higher during the last 10 ns with an average of 1.5. Both palmitate and stearate also found have biological activity against several virus including adenovirus, poxvirus, and influenza virus with score greater than 0.5 (score from 0 to 1).","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86945418","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}
Pub Date : 2022-11-14DOI: 10.35118/apjmbb.2022.030.4.01
Norainfarahin Zainal Aznam, T. A. Abdul Rahman, Ruzi Hamimi Razali, Z. Ismail, Aletza Mohd Ismail, S. S. Feng, M. Singh
AMP-activated protein kinase (AMPK) is a heterotrimeric complex that is made up of α-, β- and γ- subunits isoforms making up various isomers. AMPK has been shown to improve catabolic ATP-generating pathways, making it a promising therapeutic target. Studies have shown that different AMPK isomers are predominant in different tissues. However, determination of the predominant isomer in adipose tissue remains obscure. The goal of this study is to identify the AMPK isomers present in human subcutaneous adipose tissues comparing between lean and overweight/obese subjects. Total RNA from post-mortem subcutaneous adipose tissue was extracted and its quantity and integrity were evaluated using a bioanalyzer before being reverse transcribed to cDNA. Then, by using SYBR-green as a fluorophore detection, quantitative real-time PCR was performed. The relative expression of each target gene was calculated using the comparative 2-∆∆Ct method, and the expression levels of target genes were normalized to that of GAPDH, RPLP0, and HPRT1. Mean differences of AMPK subunits mRNA level for both lean and overweight/obese subjects were determined by Mann-Whitney for two-sample comparisons using SPSS 26.0 (SPSS Inc., USA). This study revealed that in overweight/obese subjects, the gene expression of α1-AMPK was upregulated (2.5-fold) whilst α2-AMPK was downregulated (0.5-fold) when compared to lean controls (p=0.044). There were no differences observed between the β and γ subunits in both lean and overweight/obese subjects. Findings from this study suggests that α1- and α2-AMPK gene expression plays an important role in overweight/obese subjects. Future studies to explore the AMPK gene expressions in visceral adipose tissue and among underweight subjects would further close this knowledge gap.
{"title":"Differential expression of AMPK subunit isoforms in subcutaneous adipose tissue of post-mortem subjects with BMI>25kg/m2","authors":"Norainfarahin Zainal Aznam, T. A. Abdul Rahman, Ruzi Hamimi Razali, Z. Ismail, Aletza Mohd Ismail, S. S. Feng, M. Singh","doi":"10.35118/apjmbb.2022.030.4.01","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.4.01","url":null,"abstract":"AMP-activated protein kinase (AMPK) is a heterotrimeric complex that is made up of α-, β- and γ- subunits isoforms making up various isomers. AMPK has been shown to improve catabolic ATP-generating pathways, making it a promising therapeutic target. Studies have shown that different AMPK isomers are predominant in different tissues. However, determination of the predominant isomer in adipose tissue remains obscure. The goal of this study is to identify the AMPK isomers present in human subcutaneous adipose tissues comparing between lean and overweight/obese subjects. Total RNA from post-mortem subcutaneous adipose tissue was extracted and its quantity and integrity were evaluated using a bioanalyzer before being reverse transcribed to cDNA. Then, by using SYBR-green as a fluorophore detection, quantitative real-time PCR was performed. The relative expression of each target gene was calculated using the comparative 2-∆∆Ct method, and the expression levels of target genes were normalized to that of GAPDH, RPLP0, and HPRT1. Mean differences of AMPK subunits mRNA level for both lean and overweight/obese subjects were determined by Mann-Whitney for two-sample comparisons using SPSS 26.0 (SPSS Inc., USA). This study revealed that in overweight/obese subjects, the gene expression of α1-AMPK was upregulated (2.5-fold) whilst α2-AMPK was downregulated (0.5-fold) when compared to lean controls (p=0.044). There were no differences observed between the β and γ subunits in both lean and overweight/obese subjects. Findings from this study suggests that α1- and α2-AMPK gene expression plays an important role in overweight/obese subjects. Future studies to explore the AMPK gene expressions in visceral adipose tissue and among underweight subjects would further close this knowledge gap.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91348644","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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