Novel cortex lytic enzymes in Bacillus megaterium QM B1551 spores.

Present models for spore germination in Bacillus species include a requirement for either the SleB or CwlJ cortex lytic enzymes to efficiently depolymerise the spore cortex. Previous work has demonstrated that B. megaterium spores may differ to other species in this regard, since sleB cwlJ null mutant spores complemented with the gene in trans for the non-peptidoglycan lysin YpeB can efficiently degrade the cortex. Here, we identify two novel cortex lytic enzymes, encoded at the BMQ_2391 and BMQ_3234 loci, which are essential for cortex hydrolysis in the absence of SleB and CwlJ. Ellipsoid localisation microscopy places the BMQ_3234 protein within the inner-spore coat, a region of the spore that is populated by other cortex lytic enzymes. The findings reinforce the idea that there is a degree of variation in mechanisms of cortex hydrolysis across the Bacillales, raising potential implications for environmental decontamination strategies based upon targeted inactivation of components of the spore germination apparatus.BAR was the recipient of a scholarship from the Sultanate of Oman government. AG was the recipient of a Cambridge Nehru Scholarship and received support from the Raymond and Beverly Sackler Foundation

The crystal structure of Clostridium perfringens SleM, a muramidase involved in cortical hydrolysis during spore germination.

Clostridium perfringens spores employ two peptidoglycan lysins to degrade the spore cortex during germination. SleC initiates cortex hydrolysis to generate cortical fragments that are degraded further by the muramidase SleM. Here, we present the crystal structure of the C. perfringens S40 SleM protein at 1.8 Å. SleM comprises an N-terminal catalytic domain that adopts an irregular α/β-barrel fold that is common to GH25 family lysozymes, plus a C-terminal fibronectin type III domain. The latter is involved in forming the SleM dimer that is evident in both the crystal structure and in solution. A truncated form of SleM that lacks the FnIII domain shows reduced activity against spore sacculi indicating that this domain may have a role in facilitating the position of substrate with respect to the enzyme's active site. Proteins 2016; 84:1681-1689. © 2016 Wiley Periodicals, Inc.Sultanate of Oman government (scholarship)This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/prot.2511

Novel cortex lytic enzymes in <i>Bacillus megaterium</i> QM B1551 spores

ABSTRACT Present models for spore germination in Bacillus species include a requirement for either the SleB or CwlJ cortex lytic enzymes to efficiently depolymerise the spore cortex. Previous work has demonstrated that B. megaterium spores may differ to other species in this regard, since sleB cwlJ null mutant spores complemented with the gene in trans for the non-peptidoglycan lysin YpeB can efficiently degrade the cortex. Here, we identify two novel cortex lytic enzymes, encoded at the BMQ_2391 and BMQ_3234 loci, which are essential for cortex hydrolysis in the absence of SleB and CwlJ. Ellipsoid localisation microscopy places the BMQ_3234 protein within the inner-spore coat, a region of the spore that is populated by other cortex lytic enzymes. The findings reinforce the idea that there is a degree of variation in mechanisms of cortex hydrolysis across the Bacillales, raising potential implications for environmental decontamination strategies based upon targeted inactivation of components of the spore germination apparatus.</jats:p

The crystal structure of <i>Clostridium perfringens</i> SleM, a muramidase involved in cortical hydrolysis during spore germination

This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/prot.25112$\textit{Clostridium perfringens}$ spores employ two peptidoglycan lysins to degrade the spore cortex during germination. SleC initiates cortex hydrolysis to generate cortical fragments that are degraded further by the muramidase SleM. Here, we present the crystal structure of the $\textit{C. perfringens}$ S40 SleM protein at 1.8 Å. SleM comprises an N-terminal catalytic domain that adopts an irregular $\alpha/\beta$-barrel fold that is common to GH25 family lysozymes, plus a C-terminal fibronectin type III domain. The latter is involved in forming the SleM dimer that is evident in both the crystal structure and in solution. A truncated form of SleM that lacks the FnIII domain shows reduced activity against spore sacculi indicating that this domain may have a role in facilitating the position of substrate with respect to the enzyme’s active site.Sultanate of Oman government (scholarship

Model-Assisted Optimization of Cobalt Biosorption on Macroalgae Padina pavonica for Wastewater Treatment

The release of heavy metals into the environment as a result of industrial and agricultural activities represents one of the century&rsquo;s most significant issues. Cobalt is a hazardous metal that is employed in a variety of industries. In this study, response surface methodology (RSM) combined with Box&ndash;Behnken design (BBD) was utilized to optimize the Co(II) ion removal from synthetic wastewater by the brown macroalga Padina pavonica. The influence of three factors, namely algal inoculum size, pH, and initial metal concentration, was assessed in optimization studies. RSM proposed a second-order quadratic model with a p-value of &lt;0.0001 and R2 of 0.984 for P. pavonica. According to the data related to RSM optimization, the maximum percentage of Co(II) removal of 84.3% was attained under the conditions of algal inoculum size of 5.98 g/L, pH of 6.73, and initial Co(II) concentration of 21.63 mg/L. The experimental data from the biosorption process were fitted well with the Langmuir, Freundlich, and Temkin isotherm models. The maximal Co(II) adsorption capacity was estimated using the Langmuir model to be 17.98 mg/g. Furthermore, the pseudo-second-order kinetic model was shown to have the best fit for Co biosorption by P. pavonica, showing that the mechanism of Co(II) biosorption was chemisorption controlled by surface biosorption and intra-particle diffusion. Thermodynamic parameters were also investigated to evaluate the Gibbs free energy for the Co(II) ion, which was positive, showing that the biosorption process is nonspontaneous and exothermic, and the cobalt biosorption rate decreases with increasing temperature. Algal biomass was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and energy dispersive spectroscopy. These analyses revealed the biosorbent&rsquo;s diverse functional groups and porous, rough appearance. Therefore, P. pavonica can be used to implement sustainable, eco-friendly, and acceptable solutions to water pollution problems