Hyposterolactone A, a 3α-Hydroxy Steroidal Lactone from the Deep-Sea-Derived Fungus Hypocrea sp. ZEN14.

Chemical investigation of the deep-sea-derived fungus Hypocrea sp. ZEN14 afforded a new 3α-hydroxy steroidal lactone, hyposterolactone A (1) and 25 known secondary metabolites (2-26). The structure of the new compound was established by detailed spectroscopic analysis, electronic circular dichroism (ECD) calculation as well as a J-based configuration analysis. Compound 10 showed potent cytotoxicity against Huh7 and Jurkat cells with IC50 values of 1.4 µM and 6.7 µM, respectively.

Metabolic tuning of a stable microbial community in the surface oligotrophic Indian Ocean revealed by integrated meta-omics.

Understanding the mechanisms, structuring microbial communities in oligotrophic ocean surface waters remains a major ecological endeavor. Functional redundancy and metabolic tuning are two mechanisms that have been proposed to shape microbial response to environmental forcing. However, little is known about their roles in the oligotrophic surface ocean due to less integrative characterization of community taxonomy and function. Here, we applied an integrated meta-omics-based approach, from genes to proteins, to investigate the microbial community of the oligotrophic northern Indian Ocean. Insignificant spatial variabilities of both genomic and proteomic compositions indicated a stable microbial community that was dominated by Prochlorococcus, Synechococcus, and SAR11. However, fine tuning of some metabolic functions that are mainly driven by salinity and temperature was observed. Intriguingly, a tuning divergence occurred between metabolic potential and activity in response to different environmental perturbations. Our results indicate that metabolic tuning is an important mechanism for sustaining the stability of microbial communities in oligotrophic oceans. In addition, integrated meta-omics provides a powerful tool to comprehensively understand microbial behavior and function in the ocean. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-021-00119-6.

Anti-HIV Compounds from the Deep-Sea-Derived Fungus Chaetomium globosum.

Chemical investigation on the deep-sea-derived fungus Chaetomium globosum led to the isolation of nine compounds. By extensive analyses of the 1D and 2D NMR as well as HR-ESI-MS spectra, their structures were elucidated as xylariol A (1), 1,3-dihydro-4,5,6-trihydroxy-7-methylisobenzofuran (2), epicoccone B (3), epicoccolide B (4), chaetoglobosin G (5), chaetoglobosin Fex (6), cochliodone A (7), cochliodone B (8), and chaetoviridin A (9), assorting as four phenolics (1-4), two cytochalosans (5-6), and three azaplilones (7-9). Compounds 1-3 were firstly reported from C. globosum. Under the concentrations of 20 µg/mL, 1, 2, and 3 exhibited potent in vitro anti-HIV activity with the inhibition rates of 70 %, 75 %, and 88 %, respectively.

Characterization of two novel psychrophilic and piezotolerant strains, Shewanella psychropiezotolerans sp. nov. and Shewanella eurypsychrophilus sp. nov, adapted to an extreme deep-sea environment.

Three marine bacterial strains designated YLB-06T, YLB-08T and YLB-09 were isolated under high hydrostatic pressure from deep-sea sediment samples collected from the Southwest Indian Ocean. They were Gram-stain-negative, oxidase- and catalase-positive, facultative anaerobic and motile. In addition, the strains were capable of growing at 0-20 °C (optimum 4-10 °C) and 0.1-40 MPa (optimum 0.1 MPa), were psychrophiles and piezotolerant, and could use trimethylamine N-oxide (TMAO), DMSO, elemental sulfur and insoluble Fe (III) as terminal electron acceptors during anaerobic growth. Strain YLB-06T could also use nitrate, and strains YLB-08T and YLB-09 could use nitrite as a terminal electron acceptor. Phylogenetic tree analyses based on 16S rRNA gene sequences and 400 optimized universal marker sequences indicated that the strains belonged to the genus Shewanella. The 16S rRNA gene highest similarity, together with the estimated ANI and DDH values for these strains with their related type strains, were below the respective thresholds for species differentiation. The ANI and DDH values between YLB-08T and YLB-09 were 99.9% and 91.8%, respectively, implying that they should belong to the same genospecies. The YLB-06T genome had duplicated genes, and multiple movement modalities, attachment modalities, biofilm synthesis systems, intercellular interactions and a strong antioxidant system, which were all beneficial for survival in an extreme deep-sea environment. The G + C contents of strains YLB-06T, YLB-08T and YLB-09 were 45.1, 43.5 and 43.6 mol%, respectively. Based on polyphasic taxonomic properties, two novel psychropiezotolerant species are proposed, Shewanella psychropiezotolerans sp. nov. with YLB-06T (=MCCC 1A12715T = KCTC 62907T) and S. eurypsychrophilus sp. nov with YLB-08T (=MCCC 1A12718T = KCTC 62909T) as type strains.

Pseudoroseicyclus tamaricis sp. nov., isolated from seashore sediment of a Tamarix chinensis forest and emended descriptions of the genus Pseudoroseicyclus Park et al. 2016.

An aerobic, Gram-stain-negative, bacterium, designated CLL3-39T was isolated from seashore sediment collected at a Tamarix chinensis forest in the Marine Ecology Special Reserve of Changyi, Shandong Province, PR China. Cells of strain CLL3-39T were olive-shaped and no flagellum was observed. Strain CLL3-39T grew optimally at 33 °C, pH 7.5 and salinity (sea salts) of 40 g l-1. The main fatty acids in the cell membrane of strain CLL3-39T comprised anteiso-C15 : 0 (22.3 %), iso-C15 : 0 (14.0 %), C16 : 0 (9.2 %) and summed feature 8 (contains C18 : 1 ω7c/C18 : 1 ω6c. 26. 7 %). The main polar lipids of CLL3-39T were phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. The respiratory quinone was Q10. The G+C content of the genomic DNA of strain CLL3-39T was 69.6 mol%. The average nucleotide identity between CLL3-39T and Pseudoroseicyclus aestuarii DB-4T was 74.7 % and the in silico DNA-DNA hybridization value was 20.1 %. Phylogenetically, strain CLL3-39T belonged to the genus Pseudoroseicyclus, branching with only one type strain P. aestuarii DB-4T with 96.3 % 16S rRNA gene similarity, followed by Limimaricola cinnabarinus LL-001T (95.2 %). Based on its phenotypic, phylogenetic and chemotaxonomic characteristics, we propose strain CLL3-39T (=MCCC 1A14815T =KCTC 72665T) as a representative of a novel species in the genus Pseudoroseicyclus, for which the name Pseudoroseicyclus tamaricis sp. nov. is proposed.

Metagenomic Analysis Reveals Microbial Community Structure and Metabolic Potential for Nitrogen Acquisition in the Oligotrophic Surface Water of the Indian Ocean.

Despite being the world's third largest ocean, the Indian Ocean is one of the least studied and understood with respect to microbial diversity as well as biogeochemical and ecological functions. In this study, we investigated the microbial community and its metabolic potential for nitrogen (N) acquisition in the oligotrophic surface waters of the Indian Ocean using a metagenomic approach. Proteobacteria and Cyanobacteria dominated the microbial community with an average 37.85 and 23.56% of relative abundance, respectively, followed by Bacteroidetes (3.73%), Actinobacteria (1.69%), Firmicutes (0.76%), Verrucomicrobia (0.36%), and Planctomycetes (0.31%). Overall, only 24.3% of functional genes were common among all sampling stations indicating a high level of gene diversity. However, the presence of 82.6% common KEGG Orthology (KOs) in all samples showed high functional redundancy across the Indian Ocean. Temperature, phosphate, silicate and pH were important environmental factors regulating the microbial distribution in the Indian Ocean. The cyanobacterial genus Prochlorococcus was abundant with an average 17.4% of relative abundance in the surface waters, and while 54 Prochlorococcus genomes were detected, 53 were grouped mainly within HLII clade. In total, 179 of 234 Prochlorococcus sequences extracted from the global ocean dataset were clustered into HL clades and exhibited less divergence, but 55 sequences of LL clades presented more divergence exhibiting different branch length. The genes encoding enzymes related to ammonia metabolism, such as urease, glutamate dehydrogenase, ammonia transporter, and nitrilase presented higher abundances than the genes involved in inorganic N assimilation in both microbial community and metagenomic Prochlorococcus population. Furthermore, genes associated with dissimilatory nitrate reduction, denitrification, nitrogen fixation, nitrification and anammox were absent in metagenome Prochlorococcus population, i.e., nitrogenase and nitrate reductase. Notably, the de novo biosynthesis pathways of six different amino acids were incomplete in the metagenomic Prochlorococcus population and Prochlorococcus genomes, suggesting compensatory uptake of these amino acids from the environment. These results reveal the features of the taxonomic and functional structure of the Indian Ocean microbiome and their adaptive strategies to ambient N deficiency in the oligotrophic ocean.

Discovery of andrastones from the deep-sea-derived Penicillium allii-sativi MCCC 3A00580 by OSMAC strategy.

Andrastones are unusual 6,6,6,5-tetracyclic meroterpenoids that are rarely found in nature. Previously, three andrastones were obtained from the rice static fermentation extract of the deep-sea-derived fungus Penicillium allii-sativi MCCC 3A00580. Inspired by one strain many compounds (OSMAC) approach, the oat static fermentation on P. allii-sativi was conducted. As a result, 14 andrastones were isolated by UV-guided isolation. The chemical structures of the nine new compounds (1-9) was established by comprehensive analysis of the NMR, MS, ECD, and X-ray crystallography and the five known ones (10-14) were assigned by comparing their NMR, MS, and OR data with those reported in literature. Compound 1 bears a novel hemiketal moiety while 2 is the first example to possess a novel tetrahydrofuran moiety via C-7 and C-15. All isolates were tested for anti-allergic bioactivity. Compound 10, 3-deacetylcitreohybridonol, significantly decreased degranulation with the IC50 value of 14.8 µM, compared to that of 92.5 µM for the positive control, loratadine. Mechanism study indicated 10 could decrease the generation of histamine and TNF-α by reducing the accumulation of Ca2+ in RBL-2H3 cells. These findings indicate andrastones could be potential to discover new anti-allergic candidate drugs.

Solimonas marina sp. nov., isolated from deep seawater of the Pacific Ocean.

A taxonomic study was carried out on strain C16B3T, which was isolated from deep seawater of the Pacific Ocean. The bacterium was Gram-stain-negative, oxidase- and catalase- positive and rod-shaped. Growth was observed at salinities of 0-8.0 % and at temperatures of 10-45 °C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain C16B3T belonged to the genus Solimonas, with the highest sequence similarity to Solimonas terrae KIS83-12T (97.2 %), followed by Solimonas variicoloris MN28T (97.0 %) and the other four species of the genus Solimonas (94.5 -96.8 %). The average nucleotide identity and estimated DNA-DNA hybridization values between strain C16B3T and the type strains of the genus Solimonas were 74.05-79.48 % and 19.5-22.5 %, respectively. The principal fatty acids (>5 %) were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c; 20.9 %), iso-C16 : 0 (14.6 %), C16 : 1 ω5c (9.4 %), iso-C12 : 0 (8.4 %), summed feature 2 (C14 : 0 3-OH/iso I-C16 : 1 and C12 : 0 aldehyde; 6.8 %) and C16 : 0 (5.5 %). The G+C content of the chromosomal DNA was 65.37 mol%. The respiratory quinone was determined to be Q-8 (100 %). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unidentified aminolipids, six unidentified phospholipids and one unidentified polar lipid. The combined genotypic and phenotypic data show that strain C16B3T represents a novel species within the genus Solimonas, for which the name Solimonas marina sp. nov. is proposed, with the type strain C16B3T (=MCCC 1A04678T=KCTC 52314T).

Marinobacter changyiensis, sp. nov., isolated from offshore sediment.

An aerobic, Gram-stain-negative bacterium, designated CLL7-20T, was isolated from a marine sediment sample from offshore of Changyi, Shandong Province, China. Cells of strain CLL7-20T were rod-shaped, motile with one or more polar flagella, and grew optimally at pH 7.0, at 28 °C and with 3 % (w/v) NaCl. The principal fatty acids of strain CLL7-20T were C16 : 0 and summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c). The main polar lipids of strain CLL7-20T were phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG) and an unidentified aminolipid (AL). Strain CLL7-20T contained Q-9 as the major respiratory quinone. The G+C content of its genomic DNA was 56.2 mol%. Phylogenetically, strain CLL7-20T branched within the genus Marinobacter, with M. daqiaonensis YCSA40T being its closest phylogenetic relative (96.7 % 16S rRNA gene sequence similarity), followed by M. sediminum R65T (96.6 %). Average nucleotide identity and in silico DNA-DNA hybridization values between strain CLL7-20T and the closest related reference strains were 73.2% and 19.8 %, respectively. On the basis of its phenotypic, phylogenetic and chemotaxonomic characteristics, we suggest that strain CLL7-20T (=MCCC 1A14855T=KCTC 72664T) is the type strain of a novel species in the genus Marinobacter, for which the name Marinobacter changyiensis sp. nov. is proposed. Based on the genomic analysis, siderophore genes were found from strain CLL7-20T, which indicate its potential as a promising alternative to chemical fertilizers in iron-limitated environments such as saline soils.

Properties and Applications of Extremozymes from Deep-Sea Extremophilic Microorganisms: A Mini Review.

The deep sea, which is defined as sea water below a depth of 1000 m, is one of the largest biomes on the Earth, and is recognised as an extreme environment due to its range of challenging physical parameters, such as pressure, salinity, temperature, chemicals and metals (such as hydrogen sulphide, copper and arsenic). For surviving in such extreme conditions, deep-sea extremophilic microorganisms employ a variety of adaptive strategies, such as the production of extremozymes, which exhibit outstanding thermal or cold adaptability, salt tolerance and/or pressure tolerance. Owing to their great stability, deep-sea extremozymes have numerous potential applications in a wide range of industries, such as the agricultural, food, chemical, pharmaceutical and biotechnological sectors. This enormous economic potential combined with recent advances in sampling and molecular and omics technologies has led to the emergence of research regarding deep-sea extremozymes and their primary applications in recent decades. In the present review, we introduced recent advances in research regarding deep-sea extremophiles and the enzymes they produce and discussed their potential industrial applications, with special emphasis on thermophilic, psychrophilic, halophilic and piezophilic enzymes.

Shewanella intestini sp. nov., isolated from the intestine of abalone, Haliotis diversicolor.

A Gram-stain-negative, rod-shaped bacterium with a singular polar flagellum, designated as strain XMDDZSB0408T, was isolated from the intestine of adult abalone, Haliotis diversicolor. Activity of oxidase was positive and catalase activity was negative. It could grow at salinities from 3 to 6 % NaCl (w/v), and pH 7-9. It had the highest sequence similarity of <96.0 % with all previously established species for the complete 16S rRNA gene (1531 bp). The results of phylogenetic analysis indicated that the strain was affiliated to the genus Shewanella and closely related to Shewanella gaetbuli TF-27T and Shewanella japonica KMM 3299T (95.8 % sequence similarity), Shewanella electrodiphila MAR441T (95.6 %), Shewanella pacifica KMM 3597T (95.4 %), Shewanella donghaensis LT17T (95.3 %) and Shewanella olleyana ACEM 9T (94.7 %). The respiratory quinones were MK-7, Q-8, Q-7, MK-8, Q-7 (H4) and Q-6. The predominant fatty acids consisted of C16:0, summed feature 3 (comprised of C16 : 1ω7c/C16 : 1ω6c), C18 : 0, summed feature 8 (comprised of C18 : 1ω7c/C18 : 1ω6c), C12 : 0 and C14 : 0. The polar lipids were identified as phosphatidylethanolamine (PE), a glycolipid (GL), a phospholipid (PL) and one unidentified lipid (L). The DNA G+C content was 41.4 mol% calculated from the draft genome sequence. On the basis of its polyphasic taxonomic properties, strain XMDDZSB0408T represented a novel species, for which the name Shewanella intestini sp. nov. was proposed, with the type strain XMDDZSB0408T (=KCTC 52125T=MCCC 1A01895T).

The Histone-Like Nucleoid Structuring Protein (H-NS) Is a Negative Regulator of the Lateral Flagellar System in the Deep-Sea Bacterium Shewanella piezotolerans WP3.

Although the histone-like nucleoid structuring protein (H-NS) is well known for its involvement in the adaptation of mesophilic bacteria, such as Escherichia coli, to cold environments and high-pressure stress, an understanding of the role of H-NS in the cold-adapted benthic microorganisms that live in the deep-sea ecosystem, which covers approximately 60% of the earth's surface, is still lacking. In this study, we characterized the function of H-NS in Shewanella piezotolerans WP3, which was isolated from West Pacific sediment at a depth of 1,914 m. Anhns gene deletion mutant (WP3Δhns) was constructed, and comparative whole-genome microarray analysis was performed. H-NS had a significant influence (fold change, >2) on the expression of a variety of WP3 genes (274 and 280 genes were upregulated and downregulated, respectively), particularly genes related to energy production and conversion. Notably, WP3Δhnsexhibited higher expression levels of lateral flagellar genes than WP3 and showed enhanced swarming motility and lateral flagellar production compared to those of WP3. The DNA gel mobility shift experiment showed that H-NS bound specifically to the promoter of lateral flagellar genes. Moreover, the high-affinity binding sequences of H-NS were identified by DNase I protection footprinting, and the results support the "binding and spreading" model for H-NS functioning. To our knowledge, this is the first attempt to characterize the function of the universal regulator H-NS in a deep-sea bacterium. Our data revealed that H-NS has a novel function as a repressor of the expression of genes related to the energy-consuming secondary flagellar system and to swarming motility.

Molecular characterization of the modular chitin binding protein Cbp50 from Bacillus thuringiensis serovar konkukian.

Bacillus thuringiensis is an insecticidal bacterium whose chitinolytic system may be exploited to improve the insecticidal system of Bt-crops. A nucleotide fragment of 1368 bp from B. thuringiensis serovar konkukian S4, containing the complete coding sequence of the chitin binding protein Cbp50, was cloned and sequenced. Analyses have shown the protein to contain a modular structure consisting of an N-terminal CBM33 domain, two copies of a fibronectin-like domain and a C-terminal chitin binding domain classified as CBM5. The Cbp50 protein was heterologously expressed in Escherichia coli, purified and assessed for chitin binding activity. A deletion mutant (CBD-N; containing only the N-terminal CBM33 domain) of Cbp50 was produced to determine the role of C-terminal domains in the binding activity of the protein. The full-length Cbp50 was shown to bind ß-chitin most efficiently followed by α-chitin, colloidal chitin and cellulose. The polysaccharide binding activity of CBD-N was drastically decreased. The data demonstrate that both the N-terminal and C-terminal domains of Cbp50 are essential for the efficient binding of chitin. The purified Cbp50 showed antifungal activity against the phytopathogenic fungus Fusarium oxysporum and the opportunistic human pathogen Aspergillus niger. This is the first report of a modular chitin binding protein in bacteria.

Molecular characterization of cold-inducible beta-galactosidase from Arthrobacter sp. ON14 isolated from Antarctica.

A psychrotrophic bacterium, Arthrobacter sp. ON14, isolated from Antarctica, was shown to exhibit a high beta-galactosidase activity at a low temperature. A genomic library of ON14 was constructed and screened for beta-galactosidase genes on functional plates containing 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) as the substrate. Two different beta-galactosidase genes, named as galA, galB, were found in ON14. Computational analyses of the genes revealed that the encoded protein GalA belongs to family 2 of glycosyl hydrolysases and is a cold-active protein, whereas GalB belongs to family 42 of glycosyl hydrolysases and is a mesophilic protein. Reverse transcription analyses revealed that the expression of galA is highly induced at a low temperature (4 degrees C) and repressed at a high temperature (28degreesC) when lactose is used as the sole carbon source. Conversely, the expression of galB is inhibited at a low temperature and induced at a high temperature. The purified GalA showed its peak activity at 15 degrees C and pH 8. The mineral ions Na+, K+, Mg2+, and Mn2+ were identified as enzyme activators, whereas Ca2+ had no influence on the enzyme activity. An enzyme stability assay revealed that the activity of GalA is significantly decreased when it is incubated at 45 degrees C for 2 h, and all its activity is lost when it is incubated at 50 degrees C.

Aeromonas caviae CB101 contains four chitinases encoded by a single gene chi1.

Aeromonas caviae CB101 secretes four chitinases (around 92, 82, 70, and 55 kDa) into the culture supernatant. A chitinase gene chi1 (92 kDa) was previously studied. To identify the genes encoding the remaining three chitinases, a cosmid library of CB101 was constructed to screen for putative chitinase genes. Nine cosmid clones were shown to contain a chitinase gene on chitin plates. Surprisingly, all the positive clones contained chi1. In parallel, we purified the 55-kDa chitinase (Chi55) from the CB101 culture supernatant by continuous DEAE-Sepharose and Mono-Q anion exchange chromatography. The N-terminal amino acid sequence of the purified chitinase exactly matched the N-terminal sequence of mature Chi1, indicating that the purified chitinase (Chi55) is a truncated form of Chi1. The N- and C-terminal domains of chi1 were cloned, expressed, and purified, separately. Western blots using anti-sera to the N- and C-terminal domains of chi1 on the chitinases of CB101 showed that the four chitinases in the culture supernatant are either chi1 or C-terminal truncations of Chi1. In addition, the CB101 chi1 null mutant showed no chitinolytic activity, while CB101 chi1 null mutant complemented by pUC19chi1 containing chi1 showed all four chitinases in gel activity assay. These data indicated that all four chitinases secreted by CB101 in the culture supernatant are the product of one chitinase gene chi1.

Arthrobacter psychrochitiniphilus sp. nov., a psychrotrophic bacterium isolated from Antarctica.

A psychrotrophic Arthrobacter strain, GP3(T), was isolated from Adélie penguin guano from Antarctica and characterized. The 16S rRNA gene sequence of the novel strain showed the highest similarity (97.8 %) with that of Arthrobacter psychrolactophilus B7(T). The novel strain showed a morphological change from rod to coccus. The growth temperature range of strain GP3(T) was 0-25 degrees C, with optimal growth at 20 degrees C. The strain grew over a salinity range of between 0 and 3 % (w/v) NaCl and the optimal pH for growth was pH 6-8. Strain GP3(T) contained anteiso-C(15 : 0) as the major fatty acid. The major menaquinone was MK-9(H(2)). The cell-wall peptidoglycan type was of the A3alpha variant. The DNA G+C content of strain GP3(T) was 58.5 mol%. Strain GP3(T) was able to hydrolyse chitin, Tween 80, starch, cellulose and lactose, but not gelatin, lecithin or urea. In addition to these physiological characteristics, DNA-DNA hybridization studies clearly differentiated strain GP3(T) from A. psychrolactophilus. The combined results of phylogenetic, physiological and chemotaxonomic studies indicated that strain GP3(T) represents a novel species of the genus Arthrobacter. The name Arthrobacter psychrochitiniphilus sp. nov. (type strain GP3(T)=JCM 13874(T)=CGMCC 1.6355(T)) is proposed in recognition of the strain's strong chitin-utilizing ability.

Role and regulation of fatty acid biosynthesis in the response of Shewanella piezotolerans WP3 to different temperatures and pressures.

Members of the genus Shewanella inhabit various environments; they are capable of synthesizing various types of low-melting-point fatty acids, including monounsaturated fatty acids (MUFA) and branched-chain fatty acids (BCFA) with and without eicosapentanoic acid (EPA). The genes involved in fatty acid synthesis in 15 whole-genome-sequenced Shewanella strains were identified and compared. A typical type II fatty acid synthesis pathway in Shewanella was constructed. A complete EPA synthesis gene cluster was found in all of the Shewanella genomes, although only a few of them were found to produce EPA. The roles and regulation of fatty acids synthesis in Shewanella were further elucidated in the Shewanella piezotolerans WP3 response to different temperatures and pressures. The EPA and BCFA contents of WP3 significantly increased when it was grown at low temperature and/or under high pressure. EPA, but not MUFA, was determined to be crucial for its growth at low temperature and high pressure. A gene cluster for a branched-chain amino acid ABC transporter (LIV-I) was found to be upregulated at low temperature. Combined approaches, including mutagenesis and an isotopic-tracer method, revealed that the LIV-I transporter played an important role in the regulation of BCFA synthesis in WP3. The LIV-I transporter was identified only in the cold-adapted Shewanella species and was assumed to supply an important strategy for Shewanella cold adaptation. This is the first time the molecular mechanism of BCFA regulation in bacteria has been elucidated.