CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics
Yogurt is a coagulated dairy product that results from the fermentation of milk and milk products by lactic acid bacteria, specifically Lactobacillus bulgaricus and Streptococcus thermophilus. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging tool in molecular bi...
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Elsevier
2024-12-01
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| Series: | Food Chemistry Advances |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772753X24001680 |
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| author | G. M. M. Anwarul Hasan Anuj Kumer Das Mohammed A. Satter |
| author_facet | G. M. M. Anwarul Hasan Anuj Kumer Das Mohammed A. Satter |
| author_sort | G. M. M. Anwarul Hasan |
| collection | DOAJ |
| description | Yogurt is a coagulated dairy product that results from the fermentation of milk and milk products by lactic acid bacteria, specifically Lactobacillus bulgaricus and Streptococcus thermophilus. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging tool in molecular biology research which can be used extensively in prokaryotes and eukaryotes. Lactate dehydrogenase (LDH) is an important enzyme of glycolytic pathway. In this research, we have edited L. bulgaricus d-lactate dehydrogenase (ldh) gene using CRISPR/Cas9 system. Genome editing was accomplished through synthesizing gDNA on the basis of PAM region of ldh gene and ligation of synthesized gDNA with pCRISPR-SacB plasmid which was later co-transformed with mutagenic donor DNA into competent cells containing pCasRed. This method combined CRISPR/Cas9 with γ Red machinery to achieve editing of ldh gene. The PCR screening and sequencing results confirmed that, base deletion has been occurred in the target region of ldh gene. Differences in several characteristics including texture, pH were observed on the yogurt samples from newly developed L. bulgaricus strains compared to normally produced yogurt. Our results suggest that, other genes of lactobacillus can be edited through CRISPR/Cas9 technology which can be extensively used for metabolic engineering. |
| format | Article |
| id | doaj-art-f06b8551e23f4da28f04cb6ea27a1e73 |
| institution | Kabale University |
| issn | 2772-753X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Food Chemistry Advances |
| spelling | doaj-art-f06b8551e23f4da28f04cb6ea27a1e732024-12-13T11:08:50ZengElsevierFood Chemistry Advances2772-753X2024-12-015100772CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristicsG. M. M. Anwarul Hasan0Anuj Kumer Das1Mohammed A. Satter2Institute of Food Science and Technology (IFST), Bangladesh Council of Scientific and Industrial Research (BCSIR), Dr. Qudrat-E-Khuda Road, Dhaka 1205, Bangladesh; Corresponding author.Hi-Tech Health Care Ltd. Banani, Dhaka 1213, BangladeshInstitute of Food Science and Technology (IFST), Bangladesh Council of Scientific and Industrial Research (BCSIR), Dr. Qudrat-E-Khuda Road, Dhaka 1205, BangladeshYogurt is a coagulated dairy product that results from the fermentation of milk and milk products by lactic acid bacteria, specifically Lactobacillus bulgaricus and Streptococcus thermophilus. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging tool in molecular biology research which can be used extensively in prokaryotes and eukaryotes. Lactate dehydrogenase (LDH) is an important enzyme of glycolytic pathway. In this research, we have edited L. bulgaricus d-lactate dehydrogenase (ldh) gene using CRISPR/Cas9 system. Genome editing was accomplished through synthesizing gDNA on the basis of PAM region of ldh gene and ligation of synthesized gDNA with pCRISPR-SacB plasmid which was later co-transformed with mutagenic donor DNA into competent cells containing pCasRed. This method combined CRISPR/Cas9 with γ Red machinery to achieve editing of ldh gene. The PCR screening and sequencing results confirmed that, base deletion has been occurred in the target region of ldh gene. Differences in several characteristics including texture, pH were observed on the yogurt samples from newly developed L. bulgaricus strains compared to normally produced yogurt. Our results suggest that, other genes of lactobacillus can be edited through CRISPR/Cas9 technology which can be extensively used for metabolic engineering.http://www.sciencedirect.com/science/article/pii/S2772753X24001680LactobacillusCRISPR/Cas9Gene editingMetabolic engineering |
| spellingShingle | G. M. M. Anwarul Hasan Anuj Kumer Das Mohammed A. Satter CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics Food Chemistry Advances Lactobacillus CRISPR/Cas9 Gene editing Metabolic engineering |
| title | CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics |
| title_full | CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics |
| title_fullStr | CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics |
| title_full_unstemmed | CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics |
| title_short | CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics |
| title_sort | crispr cas9 mediated gene editing of l bulgaricus d lactate dehydrogenase ldh gene to monitor yogurt characteristics |
| topic | Lactobacillus CRISPR/Cas9 Gene editing Metabolic engineering |
| url | http://www.sciencedirect.com/science/article/pii/S2772753X24001680 |
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