New insights into the effectiveness of alpha-amylase enzyme presentation on the Bacillus subtilis spore surface by adsorption and covalent immobilization

ABSTRACT We report evidence that the CotC polypeptide, a previously identified component of the Bacillus subtilis spore coat, is assembled into at least four distinct forms. Two of these, having molecular masses of 12 and 21 kDa, appeared 8 h after the onset of sporulation and were probably assembled on the forming spore immediately after their synthesis, since no accumulation of either of them was detected in the mother cell compartment, where their synthesis occurs. The other two components, 12.5 and 30 kDa, were generated 2 h later and were probably the products of posttranslational modifications of the two early forms occurring directly on the coat surface during spore maturation. None of the CotC forms was found either on the spore coat or in the mother cell compartment of a cotH mutant. This indicates that CotH serves a dual role of stabilizing the early forms of CotC and promoting the assembly of both early and late forms on the spore surface.

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Use of IPTG-inducible promoters for anchoring recombinant proteins on the Bacillus subtilis spore surface

Protein Expression and Purification ◽

10.1016/j.pep.2013.11.018 ◽

2014 ◽

Vol 95 ◽

pp. 67-76 ◽

Cited By ~ 9

Author(s):

Quynh Anh Nguyen ◽

Wolfgang Schumann

Keyword(s):

Bacillus Subtilis ◽

Recombinant Proteins ◽

Spore Surface ◽

Subtilis Spore ◽

Inducible Promoters ◽

Bacillus Subtilis Spore

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Bacillus subtilis Spore Coat

Microbiology and Molecular Biology Reviews ◽

10.1128/mmbr.63.1.1-20.1999 ◽

1999 ◽

Vol 63 (1) ◽

pp. 1-20 ◽

Cited By ~ 370

Author(s):

Adam Driks

Keyword(s):

Bacillus Subtilis ◽

Posttranslational Processing ◽

Spore Coat ◽

Coat Proteins ◽

Spore Surface ◽

Subtilis Spore ◽

Cell Type ◽

Dormant Cell ◽

Bacillus Subtilis Spore ◽

Specialized Program

SUMMARY In response to starvation, bacilli and clostridia undergo a specialized program of development that results in the production of a highly resistant dormant cell type known as the spore. A proteinacious shell, called the coat, encases the spore and plays a major role in spore survival. The coat is composed of over 25 polypeptide species, organized into several morphologically distinct layers. The mechanisms that guide coat assembly have been largely unknown until recently. We now know that proper formation of the coat relies on the genetic program that guides the synthesis of spore components during development as well as on morphogenetic proteins dedicated to coat assembly. Over 20 structural and morphogenetic genes have been cloned. In this review, we consider the contributions of the known coat and morphogenetic proteins to coat function and assembly. We present a model that describes how morphogenetic proteins direct coat assembly to the specific subcellular site of the nascent spore surface and how they establish the coat layers. We also discuss the importance of posttranslational processing of coat proteins in coat morphogenesis. Finally, we review some of the major outstanding questions in the field.

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