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Chemolithoautotrophic Campylobacterota are widespread and predominant in worldwide hydrothermal vents, and they are key players in the turnover of zero-valence sulfur. However, at present, the mechanism of cyclooctasulfur activation and catabolism in Campylobacterota bacteria is not clearly understood. Here, we investigated these processes in a hydrothermal vent isolate named Sulfurovum indicum ST-419. A transcriptome analysis revealed that multiple genes related to biofilm formation were highly expressed during both sulfur oxidation and reduction. Additionally, biofilms containing cells and EPS coated on sulfur particles were observed by SEM, suggesting that biofilm formation may be involved in S 0 activation in Sulfurovum species. Meanwhile, several genes encoding the outer membrane proteins of OprD family were also highly expressed, and among them, gene IMZ28_RS00565 exhibited significantly high expressions by 2.53- and 7.63-fold changes under both conditions, respectively, which may play a role in sulfur uptake. However, other mechanisms could be involved in sulfur activation and uptake, as experiments with dialysis bags showed that direct contact between cells and sulfur particles was not mandatory for sulfur reduction activity, whereas cell growth via sulfur oxidation did require direct contact. This indirect reaction could be ascribed to the role of H 2 S and/or other thiol-containing compounds, such as cysteine and GSH, which could be produced in the culture medium during sulfur reduction. In the periplasm, the sulfur-oxidation-multienzyme complexes soxABXY 1 Z 1 and soxCDY 2 Z 2 are likely responsible for thiosulfate oxidation and S 0 oxidation, respectively. In addition, among the four psr gene clusters encoding polysulfide reductases, only psrA 3 B 3 C 3 was significantly upregulated under the sulfur reduction condition, implying its essential role in sulfur reduction. These results expand our understanding of the interactions of Campylobacterota with the zero-valence sulfur and their adaptability to deep-sea hydrothermal environments.

The disproportionation of inorganic sulfur compounds could be widespread in natural habitats, and microorganisms could produce energy to support primary productivity through this catabolism. However, the microorganisms that carry this process out and the catabolic pathways at work remain relatively unstudied. Here, we investigated the bacterial diversity involved in sulfur disproportionation in hydrothermal plumes from Carlsberg Ridge in the northwestern Indian Ocean by enrichment cultures. A bacterial community analysis revealed that bacteria of the genera Sulfurimonas and Sulfurovum , belonging to the phylum Campylobacterota and previously having been characterized as chemolithoautotrophic sulfur oxidizers, were the most dominant members in six enrichment cultures. Subsequent bacterial isolation and physiological studies confirmed that five Sulfurimonas and Sulfurovum isolates could disproportionate thiosulfate and elemental sulfur. The ability to disproportionate sulfur was also demonstrated in several strains of Sulfurimonas and Sulfurovum that were isolated from hydrothermal vents or other natural environments. Dialysis membrane experiments showed that S 0 disproportionation did not require the direct contact of cells with bulk sulfur. A comparative genomic analysis showed that Campylobacterota strains did not contain some genes of the Dsr and rDSR pathways ( aprAB , dsrAB , dsrC , dsrMKJOP , and qmoABC ) that are involved in sulfur disproportionation in some other taxa, suggesting the existence of an unrevealed catabolic pathway for sulfur disproportionation. These findings provide evidence for the catabolic versatility of these Campylobacterota genera, which are widely distributed in chemosynthetic environments, and expand our knowledge of the microbial taxa involved in this reaction of the biogeochemical sulfur cycle in hydrothermal vent environments. IMPORTANCE The phylum Campylobacterota , notably represented by the genera Sulfurimonas and Sulfurovum , is ubiquitous and predominant in deep-sea hydrothermal systems. It is well-known to be the major chemolithoautotrophic sulfur-oxidizing group in these habitats. Herein, we show that the mesophilic predominant chemolithoautotrophs of the genera Sulfurimonas and Sulfurovum could grow via sulfur disproportionation to gain energy. This is the first report of the chemolithoautotrophic disproportionation of thiosulfate and elemental sulfur within the genera Sulfurimonas and Sulfurovum , and this comes in addition to their already known role in the chemolithoautotrophic oxidation of sulfur compounds. Sulfur disproportionation via chemolithoautotrophic Campylobacterota may represent a previously unrecognized primary production process in hydrothermal vent ecosystems.

A novel marine hydrogen- and sulfur-oxidizing bacterium, designated strain S2-6  T , was isolated from the deep-sea sediment samples at the Longqi hydrothermal system, southwestern Indian Ocean. Cells were Gram-stain-negative, motile, short rods with a single polar flagellum. Growth was observed at 10-45 °C (optimum 33 °C), pH 5.0-8.0 (optimum pH 7.0) and 1.5 to 6.0% (w/v) NaCl with an optimum at 3.0% (w/v). The isolate was an obligate chemolithoautotroph capable of growth using thiosulfate, tetrathionate, elemental sulfur or sodium sulfide as the energy source, and oxygen or nitrate as the sole electron acceptor. When hydrogen was used as the energy source, strain S2-6  T could respire oxygen, nitrate or element sulfur. The major cellular fatty acids of strain S2-6  T were summed feature 3 (C 16:1 ω7c and/or C 16:1 ω6c), C 16:0 and summed feature 8 (C 18:1 ω7c and/or C 18:1 ω6c). The total size of its genome was 2,320,257 bp and the genomic DNA G + C content was 37.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences and core genes showed that the novel isolate belonged to the genus Sulfurimonas and was most closely related to Sulfurimonas paralvinellae GO25 T (96.8% sequence identity) and Sulfurimonas autotrophica OK10 T (95.8% sequence identity). The average nucleotide identity and DNA-DNA hybridization values between strain S2-6  T and S. paralvinellae GO25 T and S. autotrophica OK10 T were 74.6%-81.2% and 19.1%-24.6%, respectively. Based on the polyphase taxonomical data, strain S2-6  T represents a novel species of the genus Sulfurimonas, for which the name Sulfurimonas sediminis sp. nov. is proposed, with the type strain S2-6  T (= MCCC 1A14513 T  = KCTC 15854  T ).

Previously, AoamyA, the alpha amylase-encoding gene from Aspergillus oryzae, was heterologously expressed in Monascus ruber CICC41233 to promote starch hydrolysis and increase the production of Monascus pigments. The target of this study is to screen the effective alpha-amylases from M. ruber for starch fast degradation and investigated for Monascus pigments production. The 13 types of predicted alpha-amylases in the M. ruber NRRL1597 genome were divided into four classes based on EC number and into five groups based on the glycoside hydrolase sub-family. The predicted alpha-amylases MrAMY1 (protein ID 440333) and MrAMY2 (protein ID 324551) showed the closest match with AOamyA by phylogenetic analysis. The genes encoding alpha-amylase, Mramy1 and Mramy2, were cloned from M. ruber CICC41233. However, the gene sequence of Mramy1 from M. ruber CICC41233 differed from that of M.ruber NRRL1597 in the length of the intron sequence. Furthermore, the Mramy1- overexpressed strain M.ruber 440333-6A completely degraded the starch of rice grain in 2 d; in contrast, starch (40.32 mg/mL) remained when rice grain was incubated with the Mramy2- overexpressed strain, M. ruber 324551-D even after 2 d, while 45.43 mg/mL and 10.48 mg/mL starch remained after 2 d and 6 d, respectively, in wild type M. ruber CICC41233. Compared to that of M. ruber CICC41233, the total Monascus pigments and ethanol-soluble pigments in M.ruber 440333-6A increased by 71.69% and 119.33% after 6d, respectively; however, it decreased by 21.40%and 26.58% after 6d, respectively, in M. ruber 324551-D. This study demonstrated that alpha-amylase MrAMY1 was superior to MrAMY2, as it effectively degraded the starch of rice grain and enhanced Monascus pigments production., Competing Interests: Conflict of interestThe authors declare that they have no competing interests., (© King Abdulaziz City for Science and Technology 2020.)

A novel mesophilic, hydrogen-, and sulfur-oxidizing bacterium, designated strain ST-419 T , was isolated from a deep-sea hydrothermal vent plume on the Carlsberg Ridge of the Northwestern Indian Ocean. The isolate was a Gram-staining-negative, non-motile and coccoid to oval-shaped bacterium. Growth was observed at 4-50 °C (optimum 37 °C), pH 5.0-8.6 (optimum pH 6.0) and 1.0-5.0 % (w/v) NaCl (optimum 3.0 %). ST-419 T could grow chemlithoautotrophically with molecular hydrogen, sulfide, elemental sulfur and thiosulfate as energy sources. Molecular oxygen, nitrate and elemental sulfur could be used as electron acceptors. The predominant fatty acids were C 16 : 1 ω7 c , C 18 : 1 ω7 c and C 16 : 0 . The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The respiratory quinone was menaquinone MK-6 and the G+C content of the genomic DNA was 42.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that ST-419 T represented a member of genus Sulfurovum and was most closely related to Sulfurovum riftiae 1812E T , with 97.6 % sequence similarity. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between ST-419 T and S. riftiae 1812E T were 74.6 and 19.6 %, respectively. The combined genotypic and phenotypic data indicate that ST-419 T represents a novel species within the genus Sulfurovum , for which the name Sulfurovum indicum sp. nov. is proposed. The type strain is ST-419 T (=MCCC 1A17954 T =KCTC 25164 T ).

To investigate the relationship between starch hydrolysis and Monascus pigments (MPs) production, the α-amylase gene ( AOamyA ) from Aspergillus oryzae was heterologously expressed in Monascus ruber CICC41233, and we obtained a positive transformant named Monascus ruber Amy9. In M. ruber Amy9, the α-amylase activities were 6.65- and 4.26-fold higher at 72 h and 144 h, respectively, than those in the parent strain with the glucose as solo carbon medium. Surprisingly, in the MPs fermentation medium with rice powder as solo material, M. ruber Amy9 completely degraded starch at 48 h, while 43.93 and 7.29 mg/mL starch remained at 48 and 144 h, respectively, in the parent strain. Monascus ruber Amy9 accelerated starch hydrolysis, which enhanced biomass and also increased total MPs by 132% after 144 h. Compared with M. ruber CICC41233, the relative gene expression levels, as determined by a quantitative real-time polymerase chain reaction analysis, of acl2 encoding ATP-citrate lyase subunit 2, pks encoding polyketide synthase, and fasB encoding the fatty acid synthase beta subunit increased by 33.14, 145.18, and 32.15%, respectively, after 144 h in M. ruber Amy9. The up-regulated expression of these key genes in MPs synthesis contributed to the large increase in MPs production. This interesting work provided us with a new idea and a new target for the study of the MPs production., Competing Interests: Compliance with ethical standardsThe author declares that they have no conflict of interest.