Pub Date : 2022-02-15DOI: 10.7324/jabb.2022.100207
Sayed Ahmed Hanan I., Shebl Ghada, Hamza Mohamed, Haider Asraf
Phenylalanine hydroxylase (PAH) enzyme plays a key role in dopamine biosynthesis in humans, rabbits, and most mammals. Rabbits, as an ideal model, were used to study the expression of the PAH gene under different feeding conditions of faba bean (Vicia faba L., Sakha 3). PAH genes (≈1,400 bp) in control and faba beanfed rabbits were PCR amplified, sequenced, and aligned with the reference gene (acc. no. 013672). The first 320 pb of PAH sequences representing the N-terminal of the gene was almost identical. High genetic similarity values were detected in PAH gene sequences for both control and faba bean-fed rabbits with the reference gene (99.80%). The results indicated very little sequence variations, which have no effect on the enzyme activity in both control and faba bean-fed rabbits. Real Time quantitative Polymerase Chain Reaction (RT-qPCR) analysis showed that the PAH gene was overexpressed after feeding on dry faba bean form compared with feeding on the fresh form. Furthermore, Western blotting results reflected superior PAH protein due to the direct influence of feeding on dry faba bean. In conclusion, our findings indicated the direct effect of fresh and dry faba bean diet on increasing phenylalanine amino acid in rabbits’ blood, which consequently increases the expression of PAH gene, thus improving the quality of life for humans.
{"title":"Characterization and expression of phenylalanine hydroxylase in rabbits under different feeding conditions of faba bean (Vicia faba L.)","authors":"Sayed Ahmed Hanan I., Shebl Ghada, Hamza Mohamed, Haider Asraf","doi":"10.7324/jabb.2022.100207","DOIUrl":"https://doi.org/10.7324/jabb.2022.100207","url":null,"abstract":"Phenylalanine hydroxylase (PAH) enzyme plays a key role in dopamine biosynthesis in humans, rabbits, and most mammals. Rabbits, as an ideal model, were used to study the expression of the PAH gene under different feeding conditions of faba bean (Vicia faba L., Sakha 3). PAH genes (≈1,400 bp) in control and faba beanfed rabbits were PCR amplified, sequenced, and aligned with the reference gene (acc. no. 013672). The first 320 pb of PAH sequences representing the N-terminal of the gene was almost identical. High genetic similarity values were detected in PAH gene sequences for both control and faba bean-fed rabbits with the reference gene (99.80%). The results indicated very little sequence variations, which have no effect on the enzyme activity in both control and faba bean-fed rabbits. Real Time quantitative Polymerase Chain Reaction (RT-qPCR) analysis showed that the PAH gene was overexpressed after feeding on dry faba bean form compared with feeding on the fresh form. Furthermore, Western blotting results reflected superior PAH protein due to the direct influence of feeding on dry faba bean. In conclusion, our findings indicated the direct effect of fresh and dry faba bean diet on increasing phenylalanine amino acid in rabbits’ blood, which consequently increases the expression of PAH gene, thus improving the quality of life for humans.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"54 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78050691","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 : 2022-02-15DOI: 10.7324/jabb.2022.100201
Anshu Aseem Kumar, Tayalkar Trupti, R. Anita, K. Vineet, Parmar Hamendra Singh
For the mapping of quantitative trait loci associated with a particular trait or introgression of desirable traits into a variety from donor parent through marker-assisted backcross breeding, parental polymorphism survey is the first prerequisite. In the present study, a total of 695 simple sequence repeats (SSR) markers spanning over 20 linkage groups were surveyed for parental polymorphism in 5 parental combinations comprising 7 soybean genotypes, namely NRC151, PS1476, IC275, IC574373, AVKS215, JS20-34, and JS20-98, with varying levels of phosphatidylcholine (PC), protein, and lipoxygenase-2. Five different parental combinations, namely NRC151 × IC275, PS1476 × IC275, NRC151 × IC574373, JS20-34 × AVKS215, and JS20-98 × AVKS215, exhibited 40.43%, 43.02%, 36.26%, 39.42%, and 41.29% parental polymorphisms, respectively. Polymorphic markers identified in these parental combinations can be utilized for genotyping of second filial generation (F2)/ recombinant inbred lines (RILs) population generated for the identification of genomic regions associated with PC, protein content, and for the recovery of recurrent parent genome content of respective recurrent parent for development of lipoxygenase-2 free high-yielding soybean genotypes.
{"title":"Genetic polymorphism of soybean genotypes with contrasting levels of phosphatidylcholine, protein, and lipoxygenase-2","authors":"Anshu Aseem Kumar, Tayalkar Trupti, R. Anita, K. Vineet, Parmar Hamendra Singh","doi":"10.7324/jabb.2022.100201","DOIUrl":"https://doi.org/10.7324/jabb.2022.100201","url":null,"abstract":"For the mapping of quantitative trait loci associated with a particular trait or introgression of desirable traits into a variety from donor parent through marker-assisted backcross breeding, parental polymorphism survey is the first prerequisite. In the present study, a total of 695 simple sequence repeats (SSR) markers spanning over 20 linkage groups were surveyed for parental polymorphism in 5 parental combinations comprising 7 soybean genotypes, namely NRC151, PS1476, IC275, IC574373, AVKS215, JS20-34, and JS20-98, with varying levels of phosphatidylcholine (PC), protein, and lipoxygenase-2. Five different parental combinations, namely NRC151 × IC275, PS1476 × IC275, NRC151 × IC574373, JS20-34 × AVKS215, and JS20-98 × AVKS215, exhibited 40.43%, 43.02%, 36.26%, 39.42%, and 41.29% parental polymorphisms, respectively. Polymorphic markers identified in these parental combinations can be utilized for genotyping of second filial generation (F2)/ recombinant inbred lines (RILs) population generated for the identification of genomic regions associated with PC, protein content, and for the recovery of recurrent parent genome content of respective recurrent parent for development of lipoxygenase-2 free high-yielding soybean genotypes.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81715857","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 : 2022-02-15DOI: 10.7324/jabb.2022.100214
Patil Aishwarya Rajendra, M. Ravi, Swamy K.N. Raghavendra, Archer Ann Catherine, S. Sowmya, Soans Sanya Hazel, Achar Raghu Ram
Aishwarya Rajendra Patil1, Ravi M.B.1* , Raghavendra Swamy K.N.1, Ann Catherine Archer2 , Sowmya S1, Sanya Hazel Soans2, Raghu Ram Achar3 1Department of Prosthodontics, JSS Dental College and Hospital, JSS Academy of Higher Education and Research, Mysuru, India. 2Department of Microbiology, School of Life Sciences, JSS Academy of Higher Education Research, Mysuru, India. 3Division of Biochemistry, School of Life Sciences, JSS Academy of Higher Education and Research, Mysuru, India.
{"title":"Lemongrass oil disrupts the biofilm of Candida albicans MTCC 1637T on soft denture reliners at lower concentrations compared to thyme and tea tree oils","authors":"Patil Aishwarya Rajendra, M. Ravi, Swamy K.N. Raghavendra, Archer Ann Catherine, S. Sowmya, Soans Sanya Hazel, Achar Raghu Ram","doi":"10.7324/jabb.2022.100214","DOIUrl":"https://doi.org/10.7324/jabb.2022.100214","url":null,"abstract":"Aishwarya Rajendra Patil1, Ravi M.B.1* , Raghavendra Swamy K.N.1, Ann Catherine Archer2 , Sowmya S1, Sanya Hazel Soans2, Raghu Ram Achar3 1Department of Prosthodontics, JSS Dental College and Hospital, JSS Academy of Higher Education and Research, Mysuru, India. 2Department of Microbiology, School of Life Sciences, JSS Academy of Higher Education Research, Mysuru, India. 3Division of Biochemistry, School of Life Sciences, JSS Academy of Higher Education and Research, Mysuru, India.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"95 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80652576","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 : 2022-02-15DOI: 10.7324/jabb.2022.100211
Marigoudar Jyoti Bharamgoud, Kuppan Narendra
The prevalence of airborne Cladosporium was investigated for seasonal occurrence in the internal environment (bedroom) in two adjacent localities of Bengaluru. The foremost objective was to study the fungal diversity in air by culturing it on nutrient media from April 2018 to March 2019. The incidence of airborne Cladosporium species exhibited significant deviation in the localities. The sampling exhibited the highest number of Cladosporium species in the winter. Cladosporium oxysporum showed the highest prevalence, followed by Cladosporium bruhnei, Cladosporium cladosporioides, and Cladosporium herbarum. The period with a highest value of CFUs (Colony Forming Units) concentration interrelated with seasonal distribution indoor aerofungi. The current study also involved the first time molecular characterization of a new rare strain of C. bruhnei obtained from the bedroom. Using 18S rDNA sequencing, the new strain (LP3CO) was identified. The sequence was submitted to the GenBank and an accession number was established. The major fungal species identified from the bedrooms were associated with the results obtained from fungal allergen detection tests carried out on respiratory allergy patients in a major hospital in Bengaluru.
{"title":"Report of the incidence of a new rare strain of Cladosporium species and incidence of three other Cladosporium species in the intramural environment of Bengaluru, India","authors":"Marigoudar Jyoti Bharamgoud, Kuppan Narendra","doi":"10.7324/jabb.2022.100211","DOIUrl":"https://doi.org/10.7324/jabb.2022.100211","url":null,"abstract":"The prevalence of airborne Cladosporium was investigated for seasonal occurrence in the internal environment (bedroom) in two adjacent localities of Bengaluru. The foremost objective was to study the fungal diversity in air by culturing it on nutrient media from April 2018 to March 2019. The incidence of airborne Cladosporium species exhibited significant deviation in the localities. The sampling exhibited the highest number of Cladosporium species in the winter. Cladosporium oxysporum showed the highest prevalence, followed by Cladosporium bruhnei, Cladosporium cladosporioides, and Cladosporium herbarum. The period with a highest value of CFUs (Colony Forming Units) concentration interrelated with seasonal distribution indoor aerofungi. The current study also involved the first time molecular characterization of a new rare strain of C. bruhnei obtained from the bedroom. Using 18S rDNA sequencing, the new strain (LP3CO) was identified. The sequence was submitted to the GenBank and an accession number was established. The major fungal species identified from the bedrooms were associated with the results obtained from fungal allergen detection tests carried out on respiratory allergy patients in a major hospital in Bengaluru.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89540778","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 : 2022-02-15DOI: 10.7324/jabb.2022.100222
Singh Shatrupa, S. Madhulika, Bisht Sanskriti, Sharma Jai Gopal
Leaf senescence is a crucial developing phase that requires the orderly disassembly of macromolecules in order to transport the nutrients from leaves into other organs and is life-threatening for plants capability. The leaf senescence is the result of a multifaceted and highly regulated mechanism involving the corresponding activities of several pathways. A lot of progress has been made recently in understanding signaling pathways of senescence, as well as how to complete the orderly process of degeneration. This paper mainly covers recent developments in the senescence of leaf and describes the function of phytohormones and essential elements from the molecular network dynamics.
{"title":"Leaf senescence and its regulation with phytohormones and essential elements: An overview","authors":"Singh Shatrupa, S. Madhulika, Bisht Sanskriti, Sharma Jai Gopal","doi":"10.7324/jabb.2022.100222","DOIUrl":"https://doi.org/10.7324/jabb.2022.100222","url":null,"abstract":"Leaf senescence is a crucial developing phase that requires the orderly disassembly of macromolecules in order to transport the nutrients from leaves into other organs and is life-threatening for plants capability. The leaf senescence is the result of a multifaceted and highly regulated mechanism involving the corresponding activities of several pathways. A lot of progress has been made recently in understanding signaling pathways of senescence, as well as how to complete the orderly process of degeneration. This paper mainly covers recent developments in the senescence of leaf and describes the function of phytohormones and essential elements from the molecular network dynamics.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"47 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87065441","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 : 2022-02-15DOI: 10.7324/jabb.2022.100217
C. Priya, Janmeda Pracheta
This study aims to investigate the phytochemical content and antioxidant activity of the leaf, stem, latex, and bark of Euphorbia neriifolia (EN) Linn. using the solvent extraction method with petroleum ether, benzene, chloroform, ethyl acetate, methanol, and aqueous extracts. Total tannin content, total saponin content, total flavonoid content, total phenol content, and total flavonol content were investigated using spectrophotometric equivalents of the standards, tannic acid, quillaja, quercetin, gallic acid, and rutin, respectively. The EN extracts of various parts were screened for potential antioxidant activities by hydrogen peroxide scavenging assay (H2O2), ferric ion reducing antioxidant power assay (FRAP), 2,2-diphenylpicrylhydrazyl assay (DPPH), metal chelation assay, nitric oxide (NO), and superoxide (SO) scavenging methods. The quantitative analysis of phytochemicals from EN revealed the presence of tannins, saponins, flavonoids, phenols, and flavonols in considerable amounts. The in vitro antioxidant assay of EN determined that the leaf, stem, latex, and bark have prominent antioxidant potential. The results showed that all the plant parts possessed antioxidant properties which were strongly correlated with the phytoconstituents. From the present study, it can be concluded that the mean content of phytochemicals in the case of EN leaf is greater than the stem, latex, and bark of the plant and this may have contributed to its great antioxidant properties. This may also justify the frequent use of the leaf more than the stem, latex, and bark in the traditional medicinal systems for the cure of bronchial infections, abdominal swellings, inflammation, pain, and tumor.
{"title":"Quantification of phytochemicals and in vitro antioxidant activities from various parts of Euphorbia neriifolia Linn.","authors":"C. Priya, Janmeda Pracheta","doi":"10.7324/jabb.2022.100217","DOIUrl":"https://doi.org/10.7324/jabb.2022.100217","url":null,"abstract":"This study aims to investigate the phytochemical content and antioxidant activity of the leaf, stem, latex, and bark of Euphorbia neriifolia (EN) Linn. using the solvent extraction method with petroleum ether, benzene, chloroform, ethyl acetate, methanol, and aqueous extracts. Total tannin content, total saponin content, total flavonoid content, total phenol content, and total flavonol content were investigated using spectrophotometric equivalents of the standards, tannic acid, quillaja, quercetin, gallic acid, and rutin, respectively. The EN extracts of various parts were screened for potential antioxidant activities by hydrogen peroxide scavenging assay (H2O2), ferric ion reducing antioxidant power assay (FRAP), 2,2-diphenylpicrylhydrazyl assay (DPPH), metal chelation assay, nitric oxide (NO), and superoxide (SO) scavenging methods. The quantitative analysis of phytochemicals from EN revealed the presence of tannins, saponins, flavonoids, phenols, and flavonols in considerable amounts. The in vitro antioxidant assay of EN determined that the leaf, stem, latex, and bark have prominent antioxidant potential. The results showed that all the plant parts possessed antioxidant properties which were strongly correlated with the phytoconstituents. From the present study, it can be concluded that the mean content of phytochemicals in the case of EN leaf is greater than the stem, latex, and bark of the plant and this may have contributed to its great antioxidant properties. This may also justify the frequent use of the leaf more than the stem, latex, and bark in the traditional medicinal systems for the cure of bronchial infections, abdominal swellings, inflammation, pain, and tumor.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88817924","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}
Thanasak Lomthong1* , Sirirat Siripornvisal2, Pannida Khunnamwong3,4 1Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Pathumthani, Thailand. 2Department of Microbiology, Faculty of Science and Technology, Phranakhon Si Ayutthaya Rajabhat University, Ayutthaya, Thailand. 3Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok, Thailand. 4Biodiversity Center Kasetsart University (BDCKU), Bangkok, Thailand
Thanasak Lomthong1*， Sirirat Siripornvisal2, Pannida khunnamwong3,4 1泰国巴吞他尼Thanyaburi拉贾曼加拉理工大学科学技术学院生物学部2泰国大城府Phranakhon Si Ayutthaya拉贾曼加拉大学科学技术学院微生物学部3泰国曼谷Kasetsart大学理学院微生物学部4泰国曼谷Kasetsart大学生物多样性中心(BDCKU
{"title":"Ultrasound-assisted enzymatic hydrolysis of broken Riceberry rice for sugar syrup production as a substrate for bacterial cellulose facial mask development","authors":"Lomthong Thanasak, Siripornvisal Sirirat, Khunnamwong Pannida","doi":"10.7324/jabb.2022.100212","DOIUrl":"https://doi.org/10.7324/jabb.2022.100212","url":null,"abstract":"Thanasak Lomthong1* , Sirirat Siripornvisal2, Pannida Khunnamwong3,4 1Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Pathumthani, Thailand. 2Department of Microbiology, Faculty of Science and Technology, Phranakhon Si Ayutthaya Rajabhat University, Ayutthaya, Thailand. 3Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok, Thailand. 4Biodiversity Center Kasetsart University (BDCKU), Bangkok, Thailand","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"71 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82924130","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}
A split-plot design field trial was carried out on Ocimum basilicum L. plants at Siwa Oasis, Egypt, during the 2018 and 2019 seasons to study the effect of organic fertilization, cultivation of some foreign varieties, and their combination on yield. The main plots included applying two compost manure levels before planting (24 and 48 m3/hectare). Subplots included cultivating five basil varieties (local and four foreign varieties: lime basil, smuglyanka basil, purple basil, and purple ruffles basil). The results revealed that applying compost manure at the highest level significantly increased herb and oil yield attributes over the lowest level. The evaluation of varieties showed that the purple ruffles and purple varieties were the most tolerant to abiotic stresses and recorded the significantly maximum herbage and oil yield. The essential oil chemical composition was affected by the interaction within treatments and harvest time. Applying 48 m3 compost/hectare and cultivation of foreign purple variety should be recommended in this region to produce the best yield quantity and quality characteristics (oil of high linalool of 50.95%–75.01% and minimum estragole of 0.00%–0.02%, as well as attractive flavor and color) which could promote exports.
{"title":"Effect of organic fertilization on productivity of some newly introduced basil varieties under Siwa Oasis conditions","authors":"Toaima Wael Ismail Mohamed, Badawy Moustafa Yehia Mohemed Ali, Hamed Emad Saleh","doi":"10.7324/jabb.2022.100210","DOIUrl":"https://doi.org/10.7324/jabb.2022.100210","url":null,"abstract":"A split-plot design field trial was carried out on Ocimum basilicum L. plants at Siwa Oasis, Egypt, during the 2018 and 2019 seasons to study the effect of organic fertilization, cultivation of some foreign varieties, and their combination on yield. The main plots included applying two compost manure levels before planting (24 and 48 m3/hectare). Subplots included cultivating five basil varieties (local and four foreign varieties: lime basil, smuglyanka basil, purple basil, and purple ruffles basil). The results revealed that applying compost manure at the highest level significantly increased herb and oil yield attributes over the lowest level. The evaluation of varieties showed that the purple ruffles and purple varieties were the most tolerant to abiotic stresses and recorded the significantly maximum herbage and oil yield. The essential oil chemical composition was affected by the interaction within treatments and harvest time. Applying 48 m3 compost/hectare and cultivation of foreign purple variety should be recommended in this region to produce the best yield quantity and quality characteristics (oil of high linalool of 50.95%–75.01% and minimum estragole of 0.00%–0.02%, as well as attractive flavor and color) which could promote exports.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"18 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84773642","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 : 2022-02-15DOI: 10.7324/jabb.2022.100215
Bhat Sagar Krishna, P. Kavya, Appu Rao Appu Rao Gopala Rao, Kini K Ramachandra
GH11 xylanase from Bacillus pumilus is a monofunctional low molecular weight single polypeptide chain xylanase with biotechnological applications. The codon-optimized xynA gene was modified to replace the signaling factor with Pichia expression system. 876 bp gene was synthesized and expressed in methylotrophic yeast Pichia pastoris GS115. The recombinant protein was purified to apparent homogeneity by conventional protein purifications methods. The specific activity, kinetic parameters Km and Vmax, temperature optima, and pH optima of the recombinant protein were similar to the native protein. The kinetic properties of native and recombinant protein indicated structural similarities of the two. The temperature stability of the recombinant protein was higher than the native protein. The half-life (t1⁄2) at 55°C was 10.5 and 21 minutes for native and recombinant xylanase, respectively. The native GH11 xylanase had a molecular weight of 22 kDa and the recombinant protein had a higher molecular weight of 25 kDa, which could be due to glycosylation. This cloning and expression of the GH11 xylanase in P. pastoris opens the possibilities of 1) production of GH11 xylanases for industrial applications in an economical way 2) creating mutants for improved activity 3) creating mutants for improved thermal stability, and 4) desirable pH optima to meet the industrial requirements.
{"title":"Cloning and expression of a GH11 xylanase from Bacillus pumilus SSP-34 in Pichia pastoris GS115: Purification and characterization","authors":"Bhat Sagar Krishna, P. Kavya, Appu Rao Appu Rao Gopala Rao, Kini K Ramachandra","doi":"10.7324/jabb.2022.100215","DOIUrl":"https://doi.org/10.7324/jabb.2022.100215","url":null,"abstract":"GH11 xylanase from Bacillus pumilus is a monofunctional low molecular weight single polypeptide chain xylanase with biotechnological applications. The codon-optimized xynA gene was modified to replace the signaling factor with Pichia expression system. 876 bp gene was synthesized and expressed in methylotrophic yeast Pichia pastoris GS115. The recombinant protein was purified to apparent homogeneity by conventional protein purifications methods. The specific activity, kinetic parameters Km and Vmax, temperature optima, and pH optima of the recombinant protein were similar to the native protein. The kinetic properties of native and recombinant protein indicated structural similarities of the two. The temperature stability of the recombinant protein was higher than the native protein. The half-life (t1⁄2) at 55°C was 10.5 and 21 minutes for native and recombinant xylanase, respectively. The native GH11 xylanase had a molecular weight of 22 kDa and the recombinant protein had a higher molecular weight of 25 kDa, which could be due to glycosylation. This cloning and expression of the GH11 xylanase in P. pastoris opens the possibilities of 1) production of GH11 xylanases for industrial applications in an economical way 2) creating mutants for improved activity 3) creating mutants for improved thermal stability, and 4) desirable pH optima to meet the industrial requirements.","PeriodicalId":15032,"journal":{"name":"Journal of Applied Biology and Biotechnology","volume":"258 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76210984","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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