Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum
Abstract Background The production of recombinant proteins with proper conformation, appropriate post-translational modifications in an easily scalable and cost-effective system is challenging. Lactococcus lactis has recently been identified as an efficient Gram positive cell factory for the product...
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doaj-d0d1b6fd44a14f48bfe4271634941caf2020-11-25T00:54:21ZengBMCMicrobial Cell Factories1475-28592018-04-0117111310.1186/s12934-018-0902-2Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparumSusheel K. Singh0Régis Wendpayangde Tiendrebeogo1Bishwanath Kumar Chourasia2Ikhlaq Hussain Kana3Subhash Singh4Michael Theisen5Department for Congenital Disorders, Statens Serum InstitutDepartment for Congenital Disorders, Statens Serum InstitutDepartment for Congenital Disorders, Statens Serum InstitutDepartment for Congenital Disorders, Statens Serum InstitutIndian Institute of Integrative MedicineDepartment for Congenital Disorders, Statens Serum InstitutAbstract Background The production of recombinant proteins with proper conformation, appropriate post-translational modifications in an easily scalable and cost-effective system is challenging. Lactococcus lactis has recently been identified as an efficient Gram positive cell factory for the production of recombinant protein. We and others have used this expression host for the production of selected malaria vaccine candidates. The safety of this production system has been confirmed in multiple clinical trials. Here we have explored L. lactis cell factories for the production of 31 representative Plasmodium falciparum antigens with varying sizes (ranging from 9 to 90 kDa) and varying degree of predicted structural complexities including eleven antigens with multiple predicted structural disulfide bonds, those which are considered difficult-to-produce proteins. Results Of the 31 recombinant constructs attempted in the L. lactis expression system, the initial expression efficiency was 55% with 17 out of 31 recombinant gene constructs producing high levels of secreted recombinant protein. The majority of the constructs which failed to produce a recombinant protein were found to consist of multiple intra-molecular disulfide-bonds. We found that these disulfide-rich constructs could be produced in high yields when genetically fused to an intrinsically disorder protein domain (GLURP-R0). By exploiting the distinct biophysical and structural properties of the intrinsically disordered protein region we developed a simple heat-based strategy for fast purification of the disulfide-rich protein domains in yields ranging from 1 to 40 mg/l. Conclusions A novel procedure for the production and purification of disulfide-rich recombinant proteins in L. lactis is described.http://link.springer.com/article/10.1186/s12934-018-0902-2Plasmodium falciparumMalariaDisulfide-rich proteinMerozoite antigensLactococcus lactis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Susheel K. Singh Régis Wendpayangde Tiendrebeogo Bishwanath Kumar Chourasia Ikhlaq Hussain Kana Subhash Singh Michael Theisen |
spellingShingle |
Susheel K. Singh Régis Wendpayangde Tiendrebeogo Bishwanath Kumar Chourasia Ikhlaq Hussain Kana Subhash Singh Michael Theisen Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum Microbial Cell Factories Plasmodium falciparum Malaria Disulfide-rich protein Merozoite antigens Lactococcus lactis |
author_facet |
Susheel K. Singh Régis Wendpayangde Tiendrebeogo Bishwanath Kumar Chourasia Ikhlaq Hussain Kana Subhash Singh Michael Theisen |
author_sort |
Susheel K. Singh |
title |
Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum |
title_short |
Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum |
title_full |
Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum |
title_fullStr |
Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum |
title_full_unstemmed |
Lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from Plasmodium falciparum |
title_sort |
lactococcus lactis provides an efficient platform for production of disulfide-rich recombinant proteins from plasmodium falciparum |
publisher |
BMC |
series |
Microbial Cell Factories |
issn |
1475-2859 |
publishDate |
2018-04-01 |
description |
Abstract Background The production of recombinant proteins with proper conformation, appropriate post-translational modifications in an easily scalable and cost-effective system is challenging. Lactococcus lactis has recently been identified as an efficient Gram positive cell factory for the production of recombinant protein. We and others have used this expression host for the production of selected malaria vaccine candidates. The safety of this production system has been confirmed in multiple clinical trials. Here we have explored L. lactis cell factories for the production of 31 representative Plasmodium falciparum antigens with varying sizes (ranging from 9 to 90 kDa) and varying degree of predicted structural complexities including eleven antigens with multiple predicted structural disulfide bonds, those which are considered difficult-to-produce proteins. Results Of the 31 recombinant constructs attempted in the L. lactis expression system, the initial expression efficiency was 55% with 17 out of 31 recombinant gene constructs producing high levels of secreted recombinant protein. The majority of the constructs which failed to produce a recombinant protein were found to consist of multiple intra-molecular disulfide-bonds. We found that these disulfide-rich constructs could be produced in high yields when genetically fused to an intrinsically disorder protein domain (GLURP-R0). By exploiting the distinct biophysical and structural properties of the intrinsically disordered protein region we developed a simple heat-based strategy for fast purification of the disulfide-rich protein domains in yields ranging from 1 to 40 mg/l. Conclusions A novel procedure for the production and purification of disulfide-rich recombinant proteins in L. lactis is described. |
topic |
Plasmodium falciparum Malaria Disulfide-rich protein Merozoite antigens Lactococcus lactis |
url |
http://link.springer.com/article/10.1186/s12934-018-0902-2 |
work_keys_str_mv |
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