Effect of Signal Peptides on the Secretion of β-Cyclodextrin Glucanotransferase in &lt;b&gt;&lt;i&gt;Lactococcus lactis&lt;/i&gt;&lt;/b&gt; NZ9000

Subramaniam, Menaga; Baradaran, Ali; Rosli, Illias; Mohamad, Rosfarizan; Yusoff, Khatijah; Raha, Abdul Rahim

doi:10.1159/000343921

Cited by 16 publications

(10 citation statements)

References 70 publications

(27 reference statements)

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“…More recently, we have isolated a novel signal peptide, SPK1 from Pediococcus pentasaceus , with the ability to secrete heterologous proteins with efficiencies comparable to Usp45 in L. lactis [25]. When SPK1 was used to secrete β-cyclodextrin glucanotransferase, although secretion efficiency was higher than USP45, total yield was found to be lower [26], thus demonstrating the complex effects brought upon by SPs, not only on the secretion of heterologous proteins, but on total protein yield as well.…”

Section: The Lactococcal Molecular Toolboxmentioning

confidence: 99%

A review on Lactococcus lactis: from food to factory

et al. 2017

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Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Salmonella cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

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Section: The Lactococcal Molecular Toolboxmentioning

confidence: 99%

A review on Lactococcus lactis: from food to factory

et al. 2017

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“…Cano‐Garrido et al () reported the solubility of aggregation‐prone GFP variant (VP1GFP), expressed using pNZ8148 in L. lactis , was influenced by time, culture conditions and growth temperature, which have a dramatic effect not only on protein yield but also on protein solubility and conformational quality that were particularly favoured under fermentative anaerobic metabolism. Despite testing an extended duration of expression (6–12 h) at different growth temperatures (including the optimum 30°C), as previously reported for secretion of recombinant proteins containing USP45 (Subramaniam et al ; Siaw et al ), the expressed recombinant S‐Ara2.02‐H remained insoluble and intracellular even after 24 h of expression. Subsequent purification and refolding of the insoluble rS‐Ara2.02‐H recombinant protein produced in L. lactis was performed to verify its identity and allergenicity prior to prophylactic study.…”

Section: Discussionmentioning

confidence: 82%

Lactococcus lactisharbouring Ara h 2.02 alleviates allergen‐specific Th2‐associated responses in sensitized mice

et al. 2019

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Aim To study the prophylactic effect of recombinant Lactococcus lactis (rLl) harbouring Ara h 2.02 peanut allergen, in sensitized and challenged mice. Methods and Results Ara h 2.02 cDNA was cloned into pNZ8048 for heterologous expression in L. lactis. The purified recombinant allergen showed IgE binding comparable with native Ara h 2. Balb/c mice were fed with either recombinant (rLl), nonrecombinant L. lactis (Ll) or NaHCO3 (Sham) prior to sensitization and challenged with rAra h 2.02, whereas the baseline group was only fed with Ll. Allergen‐specific immunoglobulin and splenocyte cytokines responses were determined for each mouse. Mice fed with either Ll or rLl showed significant alleviation of IgE and IgG1 compared to the Sham group. Despite no significant decrease in Th2 (IL‐4, IL‐13, IL‐6) or increase in Th1 (IFN‐γ) cytokines, both groups showed lower IL‐10 level, while the IL‐4 : IFN‐γ ratio was significantly lower for rLl compared to Ll group. Conclusions Oral administration of rLl harbouring Ara h 2.02 demonstrated alleviation of Th2‐associated responses in allergen‐challenged mice and a possible added allergen‐specific prophylactic effect. Significance and Impact of the Study Ara h 2.02 coupled with the intrinsic properties of probiotic L. lactis as a delivery vehicle can be explored for the development of a commercially scalable vaccine.

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“…[1] Bacterial culture in the form of glycerol stock, maintained at ¡20 C, was used for inoculum preparation. For this purpose, the stock culture was thawed and streaked on M17 agar (Merck, Germany) plate followed by incubation at 30 C for 24 h. A single colony was inoculated in 10 mL of M17 broth and incubated overnight followed by a sub-culture in fresh M17 broth and incubation at the same conditions.…”

Section: Strain and Inoculum Preparationmentioning

confidence: 99%

“…The expression system used in the present study is the nisin-controlled expression (NICE) system [1,40]. Mierau et al [40] conducted a study in order to optimize the L. lactis NICE system for industrial applications.…”

Section: Artificial Neural Networkmentioning

confidence: 99%

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Cyclodextrin glycosyltransferase biosynthesis improvement by recombinantLactococcus lactisNZ:NSP:CGT: medium formulation and culture condition optimization

Amiri

Mohamad

Rahim

et al. 2015

Biotechnology & Biotechnological Equipment

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Cyclodextrin glycosyltransferase (CGTase) is a bacterial glycosyltransferase which catalyses the conversion of starch to cyclodextrin. It can be produced industrially through a fermentation process. The optimal design of the fermentation medium and conditions is critical for metabolic production and microbial growth. Optimization of CGTase biosynthesis by recombinant Lactococcus lactis NZ:NSP:CGT was performed using an artificial neural network (ANN) as the main tool in order to improve the CGTase production in the fermentation process. Three parameters, including temperature, carbon source concentration and nitrogen source concentration, were determined as significant factors for improvement of CGTase production by L. lactis NZ:NSP:CGT in the fermentation process. Soluble potato starch and yeast extract were chosen as the best carbon and nitrogen sources for higher production of CGTase by L. lactis NZ:NSP:CGT. The optimum concentration of soluble starch and yeast extract were 3.82% and 5.67% (w/v), respectively, and the optimum temperature was 20C. The final CGTase activity under optimized conditions was 22.09 U/mL, which was close enough to the predicted value of 24.17 U/mL. The obtained results are of particular interest, as CGTase is an industrially important enzyme. In this study, its production through batch fermentation was optimized in order to maximize its production. In future studies, more investigations in various modes of fermentation should be conducted in order to increase the CGTase biosynthesis.

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Effect of Signal Peptides on the Secretion of β-Cyclodextrin Glucanotransferase in <b><i>Lactococcus lactis</i></b> NZ9000

Cited by 16 publications

References 70 publications

A review on Lactococcus lactis: from food to factory

A review on Lactococcus lactis: from food to factory

Lactococcus lactisharbouring Ara h 2.02 alleviates allergen‐specific Th2‐associated responses in sensitized mice

Cyclodextrin glycosyltransferase biosynthesis improvement by recombinantLactococcus lactisNZ:NSP:CGT: medium formulation and culture condition optimization

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