Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge

In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Mitigation of Commercial Food Waste-Related Salinity Stress Using Halotolerant Rhizobacteria in Chinese Cabbage Plants

The use of commercial food waste in the Korean agricultural industry is increasing due to its capacity to act as an ecofriendly fertilizer. However, the high salt content of food waste can be detrimental to plant health and increase salinity levels in agricultural fields. In the current study, we introduced halotolerant rhizobacteria to neutralize the negative impact of food waste-related salt stress on crop productivity. We isolated halotolerant rhizobacteria from plants at Pohang beach, and screened bacterial isolates for their plant growth-promoting traits and salt stress-mitigating capacity; consequently, the bacterial isolate Bacillus pumilus MAK9 was selected for further investigation. This isolate showed higher salt stress tolerance and produced indole-3-acetic acid along with other organic acids. Furthermore, the inoculation of B. pumilus MAK9 into Chinese cabbage plants alleviated the effects of salt stress and enhanced plant growth parameters, i.e., it increased shoot length (32%), root length (41%), fresh weight (18%), dry weight (35%), and chlorophyll content (13%) compared with such measurements in plants treated with food waste only (control). Moreover, relative to control plants, inoculated plants showed significantly decreased abscisic acid content (2-fold) and increased salicylic acid content (11.70%). Bacillus pumilus MAK9-inoculated Chinese cabbage plants also showed a significant decrease in glutathione (11%), polyphenol oxidase (17%), and superoxide anions (18%), but an increase in catalase (14%), peroxidase (19%), and total protein content (26%) in comparison to the levels in control plants. Inductively coupled plasma mass spectrometry analysis showed that B. pumilus MAK9-inoculated plants had higher calcium (3%), potassium (22%), and phosphorus (15%) levels, whereas sodium content (7%) declined compared with that in control plants. Similarly, increases in glucose (17%), fructose (11%), and sucrose (14%) contents were recorded in B. pumilus MAK9-inoculated plants relative to in control plants. The bacterial isolate MAK9 was confirmed as B. pumilus using 16S rRNA and phylogenetic analysis. In conclusion, the use of commercially powered food waste could be a climate-friendly agricultural practice when rhizobacteria that enhance tolerance to salinity stress are also added to plants.

An antimicrobial peptide specifically active against Listeria monocytogenes is secreted by Bacillus pumilus SF214

Background Members of the Bacillus genus produce a large variety of antimicrobial peptides including linear or cyclic lipopeptides and thiopeptides, that often have a broad spectrum of action against Gram-positive and Gram-negative bacteria. We have recently reported that SF214, a marine isolated strain of Bacillus pumilus, produces two different antimicrobials specifically active against either Staphylococcus aureus or Listeria monocytogenes. The anti-Staphylococcus molecule has been previously characterized as a pumilacidin, a nonribosomally synthesized lipopetide composed of a mixture of cyclic heptapeptides linked to fatty acids of variable length. Results Our analysis on the anti-Listeria molecule of B. pumilus SF214 indicated that it is a peptide slightly smaller than 10 kDa, produced during the exponential phase of growth, stable at a wide range of pH conditions and resistant to various chemical treatments. The peptide showed a lytic activity against growing but not resting cells of Listeria monocytogenes and appeared extremely specific being inactive also against L. innocua, a close relative of L. monocytogenes. Conclusions These findings indicate that the B. pumilus peptide is unusual with respect to other antimicrobials both for its time of synthesis and secretion and for its strict specificity against L. monocytogenes. Such specificity, together with its stability, propose this new antimicrobial as a tool for potential biotechnological applications in the fight against the dangerous food-borne pathogen L. monocytogenes.

Isolation and Characterization of Plant Growth-Promoting Compost Bacteria That Improved Physiological Characteristics in Tomato and Lettuce Seedlings

Currently, agricultural systems are inadequate to meet the demand of the population, coupled with the constant degradation of natural resources. Therefore, it is necessary to explore alternatives to increase the productivity and quality of crops with minimal environmental impact. The use of plant growth-promoting bacteria can provide solutions to some agri-environmental problems and replace or minimize conventional agricultural practices. In this study, a Bacillus pumilus strain with plant growth-promoting properties was isolated from mature compost. In vitro, the ability of Bacillus pumilus to solubilize phosphate, inhibit the growth of phytopathogenic fungi, and its effect on the germination of tomato and lettuce seeds was evaluated. In vivo, its effect on stem thickness, height, and the number of leaves of tomato and lettuce seedlings was studied. The results show that, in vitro, Bacillus pumilus solubilizes phosphate, inhibits the growth of the fungus Fusarium oxysporum, and increases the germination percentage of tomato seeds. The results, in vivo, demonstrate that the bacteria increases the stem thickness of tomato seedlings, while, in lettuce, it increases the stem thickness and the number of leaves. The outcome implies that Bacillus pumilus has properties as a plant growth promoter and can be used as a promising inoculant to enhance the growth of tomato and lettuce seedlings.

Cryo-EM Structures of Two Bacteriophage Portal Proteins Provide Insights for Antimicrobial Phage Engineering

Widespread antibiotic resistance has returned attention to bacteriophages as a means of managing bacterial pathogenesis. Synthetic biology approaches to engineer phages have demonstrated genomic editing to broaden natural host ranges, or to optimise microbicidal action. Gram positive pathogens cause serious pastoral animal and human infections that are especially lethal in newborns. Such pathogens are targeted by the obligate lytic phages of the Salasmaviridae and Guelinviridae families. These phages have relatively small ~20 kb linear protein-capped genomes and their compact organisation, relatively few structural elements, and broad host range, are appealing from a phage-engineering standpoint. In this study, we focus on portal proteins, which are core elements for the assembly of such tailed phages. The structures of dodecameric portal complexes from Salasmaviridae phage GA1, which targets Bacillus pumilus, and Guelinviridae phage phiCPV4 that infects Clostridium perfringens, were determined at resolutions of 3.3 Å and 2.9 Å, respectively. Both are found to closely resemble the related phi29 portal protein fold. However, the portal protein of phiCPV4 exhibits interesting differences in the clip domain. These structures provide new insights on structural diversity in Caudovirales portal proteins and will be essential for considerations in phage structural engineering.

Isolation of a Bacillus safensis from mine tailings in Peru, genomic characterization and characterization of its cyanide- degrading enzyme CynD

Cyanide is widely used in industry as a potent lixiviant due to its capacity to tightly bind metals. This property imparts cyanide enormous toxicity to all known organisms. Thus, industries that utilize this compound must reduce its concentration in recycled or waste waters. Physical, chemical, and biological treatments have been used for cyanide remediation; however, none of them meet all the desired characteristics: efficiency, low cost and low environmental impact. A better understanding of metabolic pathways and biochemistry of enzymes involved in cyanide degradation is a necessary step to improve cyanide bioremediation efficacy to satisfy the industry requirements. Here, we used several approaches to explore this topic. We have isolated three cyanide-degrading Bacillus strains from water in contact with mine tailings from Lima, Peru, and classified them as Bacillus safensis PER-URP-08, Bacillus licheniformis PER-URP-12, and Bacillus subtilis PER-URP-17 based on 16S rRNA gene sequencing and core genome analyses. Additionally, core genome analyses of 132 publicly available genomes of Bacillus pumilus group including B. safensis and B. altitudinis allowed us to reclassify some strains and identify two strains that did not match with any known species of the Bacillus pumilus group. We searched for possible routes of cyanide-degradation in the genomes of these three strains and identified putative B. licheniformis PER-URP-12 and B. subtilis PER-URP-17 rhodaneses and B. safensis PER-URP-08 cyanide dihydratase (CynD) sequences possibly involved cyanide degradation. We identified characteristic C-terminal residues that differentiate CynD from B. pumilus and B. safensis, and showed that, differently from CynD from B. pumilus C1, recombinant CynD from the Bacillus safensis PER-URP-08 strain remains active up to pH 9 and presents a distinct oligomerization pattern at pH 8 and 9. Moreover, transcripts of B. safensis PER-URP-08 CynD (CynDPER-URP-08) are strongly induced in the presence of cyanide. Our results warrant further investigation of B. safensis PER-URP-08 and CynDPER-URP-08 as potential tools for cyanide-bioremediation.

Long-read sequencing reveals increased occurrence of genomic variants and adenosine methylation in Bacillus pumilus SAFR-032 after long-duration flight exposure onboard the International Space Station

Bacillus pumilus SAFR-032, an endospore-forming bacterial strain, was investigated to determine its methylation pattern (methylome) change, compared to ground control, after direct exposure to space conditions onboard the International Space Station (ISS) for 1.5 years. The resulting ISS-flown and non-flown strains were sequenced using the Nanopore MinION and an in-house method and pipeline to identify methylated positions in the genome. Our analysis indicated genomic variants and m6A methylation increased in the ISS-flown SAFR-032. To complement the broader omics investigation and explore phenotypic changes, ISS-flown and non-flown strains were compared in a series of laboratory-based chamber experiments using an X-ray irradiation source (doses applied at 250, 500, 750, 1000 and 1250 Gy); results show a potentially higher survival fraction of ISS-flown DS2 at the two highest exposures. Taken together, results from this study document lasting changes to the genome by methylation, potentially triggered by conditions in spaceflight, with functional consequences for the resistance of bacteria to stressors expected on long-duration missions beyond low Earth orbit.