Jati, Afif Pranaya (2018) Identification and Production Of Novel Lantibiotics From Clostridium Species Via Heterologous Expression in Lactococcus lactis. Master's Research Project 1, Molecular Biology and Biotechnology (2016-2019).
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Abstract
The capability of nisin induced gene expression system (NICE) in Lactococcus lactis to modify small peptides into lanthipeptides, provide an exciting opportunity for novel antimicrobials production to combat various pathogens. This system can be exploited to produce “new to nature” lantibiotics from various organisms using synthetic biology approach. This study aimed to identify and produce novel lantibiotics from the genus of Clostridium that had not been discovered. As the result, 54 putative clostridia new lantibiotic genes were discovered after in silico analysis with BAGEL3 and Anti-SMASH. Subsequently, based on the novelty and the compatibility to expression system, twelve putative clostridia lantibiotic genes were selected as candidates and tested for production via heterologous expression in Lactococcus lactis. Interestingly, eleven peptides except Clos16 displayed antimicrobial activity against L. lactis NZ9000 as indicator strain. Moreover, Micrococcus flavus is susceptible to Clos4, Clos12, Clos22 and ClosDP. Additionally, Clos2, Clos14 and ClosDP lantibiotics showed antimicrobial activity against Clostridium sporogenes and Micrococcus luteus. Nevertheless, mass spectrometry analyses using MALDI-TOF and LC-MS/MS revealed multiple dehydrated serines and/or threonines in several putative lantibiotic candidates, confirming successful production and secretion of novel lantibiotics from the genus of Clostridium by nisin synthetic machinery.
| Item Type: | Thesis (Master's Research Project 1) |
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| Supervisor name: | Kuipers, O.P. |
| Degree programme: | Molecular Biology and Biotechnology (2016-2019) |
| Thesis type: | Master's Research Project 1 |
| Language: | English |
| Date Deposited: | 20 Feb 2018 |
| Last Modified: | 27 Mar 2018 13:33 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/16492 |
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