Design of signal peptide bombyxin and its effect on secretory expression efficiency and levels of Helicobacter pylori urease subunit B in silkworm cells and larvae

Zhang, Xiaolin; Jiang, Anmin; Ma, Zhongyou; Xiong, Youyi; Dou, Jing; Zhou, Guoliang; Qin, Meisong; Wang, Jianfei

doi:10.1590/s1516-8913201500013

Cited by 5 publications

(3 citation statements)

References 22 publications

(24 reference statements)

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“…To counterpart H. pylori infection in humans, an secretory system have been developed that consists of Ure B gene (Urease) constructed along with signal peptide bombyxin from B. mori. The expression system was found to be higher in terms of production and would aid in the large-scale expression, yield of UreB in silkworm larvae [79]. In fact, the oral immunization of silkworm pupae powder containing recombinant UreB of Helicobacter pylori provided therapeutic effects against Helicobacter pylori infection when tested in mice [80].…”

Section: Discussionmentioning

confidence: 99%

Detection of gut microflora and enzymes using metagenomics and metaproteomics in silkworm, Bombyx mori

Mohanraj

Mahalingam

Karthikeyan³

et al. 2023

Preprint

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Background Microorganisms living in insect gut plays a crucial role in the adaptation, growth and development of the insect hosts. The identification and molecular analysis of insect gut microbes will enable us to develop novel strategies for industrial product development, effective utilization of by products and facilitate us with the best pest management practices. In our present study, we employed a culture independent metagenomic approach in conjunction with metaproteomic profiling to enumerate the gut microbes in pure races and cross breeds of silkworm, Bombyx mori L.Results The phylogenetic analysis showed that the gut microflora was majorly grouped into four bacterial classes: Gamma Proteobacteria, Actinobacteria, Cocci and Bacilli. Two novel proteins in both silkworm races were identified and an additional three proteins were found to be differentially expressed between the races.Conclusions Silkworm undergoes radical morphological variations upon metamorphosis and a key challenge in studying gut microflora is the ability to access different genomes and the corresponding protein profiles. The results presented in this study may help to bridge the gap between the unknowns of silkworm gut microbiota and their importance in the silkworm gut ecosystem.

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Section: Discussionmentioning

confidence: 99%

Detection of gut microflora and enzymes using metagenomics and metaproteomics in silkworm, Bombyx mori

Mohanraj

Mahalingam

Karthikeyan³

et al. 2023

Preprint

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“…Bombyxin signal peptide was previously utilized to target non-secretory proteins, such as urease subunit B and β1,3-N-acetylglucosamyltransferase 2, for export via the secretion pathway of an insect expression system 14 , 15 , 22 . Secretory expression improves protein production yield, underscoring the strategy of using the bombyxin signal peptide as effective in a large-scale protein production platform 22 . Secreted proteins may be extracted from either silkworm hemolymph or fat body.…”

Section: Discussionmentioning

confidence: 99%

“…Due to broad and intricate biological activities, growth factors are increasingly recognized as valuable in research, theranostic and biopharmaceutical applications. Different approaches at manipulating intrinsic and bioengineered signal peptides have been evaluated for optimizing heterologous expression of growth factors 11 – 22 . Nevertheless, structural sequence homogeneity at the signal peptide cleavage site in bioengineered growth factors secreted by the unconventional pathway is largely undocumented.…”

Section: Introductionmentioning

confidence: 99%

Conventional and unconventional secretory proteins expressed with silkworm bombyxin signal peptide display functional fidelity

Park

Arrell

Reyes

et al. 2017

Sci Rep

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Growth factors are signaling molecules which orchestrate cell growth, proliferation and differentiation. The majority are secreted proteins, exported through the classical endoplasmic reticulum (ER)/Golgi-dependent pathway, but a few are released by unconventional ER/Golgi-independent means. Human fibroblast growth factor 2 (FGF2) and insulin-like growth factor 1 (IGF1), are canonical prototypes secreted by the unconventional and conventional pathway, respectively. We herein examined whether expression of these two growth factors in the Bombyx mori nucleopolyhedrovirus (BmNPV)-based silkworm expression system with its innate signal peptide, bombyxin, secures structural homogeneity at the signal peptide cleavage site regardless of the native secretory route. Proteomic analysis mapped structural microheterogeneity of signal peptide cleavage at the amino terminus of FGF2, whereas IGF1 displayed homogeneous amino-terminal cleavage with complete removal of the bombyxin signal peptide. A cell proliferation assay revealed potent functional activity of both FGF2 and IGF1, suggesting that FGF2 amino-terminal microheterogeneity does not alter mitogenic activity. These findings demonstrate that the occurrence of amino-terminal structural homogeneity may be associated with the original secretion mechanism of a particular growth factor. Furthermore, our results highlight the bombyxin signal peptide as a reliable secretion sequence applicable to mass production of functionally active secretory proteins in a silkworm-based expression platform.

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HIV Capsid and Protease, New Targets of Melittin

Dehghani

Hasanshahi

Hashempour

2020

Int J Pept Res Ther

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Design of signal peptide bombyxin and its effect on secretory expression efficiency and levels of Helicobacter pylori urease subunit B in silkworm cells and larvae

Abstract: This study employed a Bac-to-Bac/Bombyx mori bioreactor to mass-produce immunogenic urease subunit B (UreB

Cited by 5 publications

References 22 publications

Detection of gut microflora and enzymes using metagenomics and metaproteomics in silkworm, Bombyx mori

Detection of gut microflora and enzymes using metagenomics and metaproteomics in silkworm, Bombyx mori

Conventional and unconventional secretory proteins expressed with silkworm bombyxin signal peptide display functional fidelity

HIV Capsid and Protease, New Targets of Melittin

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