Bacterial carbonic anhydrase-induced carbonates mitigate soil erosion in biological soil crusts.

Soil erosion is a significant environmental issue worldwide, particularly in island regions where land resources are exceedingly scarce. Biological soil crusts play a crucial role in mitigating soil erosion, yet the precise effect and mechanism of biological soil crusts against erosion remain ambiguous. In this study, biological soil crusts at various developmental stages from a tropical coral island in the South China Sea were chosen to investigate the role of carbonic anhydrase in mitigating erosion. A cohesive strength meter, real-time quantitative PCR, and 16S rRNA gene high-throughput sequencing were employed to assess variations in soil antiscouribility as well as bacterial abundance and composition during the formation and development of biological soil crusts. Scanning electron microscopy was utilized to detect carbonates induced by bacterial carbonic anhydrase and elucidate their role in the solidification of sand particles. The findings indicate that the formation and development of biological soil crusts significantly enhance anti-scouribility. Comparison to those of bare coral sand, the shear stress increased from 0.35 to 1.11 N/m2 in the dark biocrusts. Moreover, significantly elevated carbonic anhydrase activity was observed in biological soil crusts, demonstrating a positive correlation with antiscouribility. In addition, there was a significant increase in bacterial abundance within the biological soil crusts. The enrichment of Cyanobacteriales and Chloroflexales potentially contributed to the increased carbonic anhydrase activity and antiscouribility. Furthermore, three cyanobacterial strains with carbonic anhydrase activity were isolated from biological soil crusts and subsequently confirmed to enhance sand solidification through microbial carbonate precipitation. This study presents initial evidence for the role of microbial carbonic anhydrase in enhancing the antiscouribility of biological soil crusts during their formation and development. These findings offer novel insights into the functional and mechanistic dimensions underlying the mitigation of soil erosion facilitated by biological soil crusts, which are valuable for implementing sustainable biorestoration and environmental management technologies to prevent soil erosion.

Characterization of a novel isoflavone glycoside-hydrolyzing ß-glucosidase from mangrove soil metagenomic library.

For the beneficial pharmacological properties of isoflavonoids and their related glycoconjugates, there is increasingly interest in their enzymatic conversion. In this study, a novel ß-glucosidase gene isolated from metagenomic library of mangrove sediment was cloned and overexpressed in Escherichia coli BL21(DE3). The purified recombination ß-glucosidase, designated as r-Bgl66, showed high catalytic activity for soy isoflavone glycosides. It converted soy isoflavone flour extract with the productivities of 0.87 mM/h for daidzein, 0.59 mM/h for genistein and 0.42 mM/h for glycitein. The kcat/Km values for daidzin, genistin and glycitin were 208.73, 222.37 and 288.07 mM-1 s-1, respectively. In addition, r-Bgl66 also exhibited the characteristic of glucose-tolerance, and the inhibition constant Ki was 471.4 mM. These properties make it a good candidate in the enzymatic hydrolysis of soy isoflavone glycosides. This study also highlights the utility of metagenomic approach in discovering novel ß-glucosidase for soy isoflavone glycosides hydrolysis.

Biodegradation and metabolic pathway of phenanthrene by a newly isolated bacterium Gordonia sp. SCSIO19801.

The bacterium Gordonia sp. SCSIO19801, which could effectively utilize phenanthrene as the sole carbon source, was isolated from the seawater of the South China Sea. Its biodegradation characteristics, whole genome sequence, and biodegradation pathway were investigated. The phenanthrene biodegradation process of Gordonia sp. SCSIO19801 was estimated to be a first-order kinetic model with a k value of 0.26/day. Based on the identification of metabolites, utilization of probable intermediates, and genomics analysis of related genes, the degradation of phenanthrene by Gordonia sp. SCSIO19801 was proposed to occur via the salicylate metabolic pathway. This is the first report of a phenanthrene degradation pathway in Gordonia species. In addition, the Gordonia sp. SCSIO19801 could use other aromatic compounds as the sole source of carbon and energy. These characteristics indicate that Gordonia sp. SCSIO19801 can be utilized for developing effective methods for the biodegradation of petroleum hydrocarbons in marine environments.

Euroticins A and B, Two Pairs of Highly Constructed Salicylaldehyde Derivative Enantiomers from a Marine-Derived Fungus Eurotium sp. SCSIO F452.

Two pairs of salicylaldehyde derivative enantiomers, euroticins A and B (1 and 2), were isolated from a marine-derived fungus Eurotium sp. SCSIO F452. Compound 1 possesses a highly constructed 6/6/6/5/7 pentacyclic structure featuring an unprecedented 2,11-dioxatricyclo[5.3.1.04,8]undecane core. Compound 2 represents the first example of 6/6/6/6 tetracyclic salicylaldehyde derivative. Their structures were established by spectroscopic analyses, X-ray diffraction, and electronic circular dichroism (ECD) and 13C NMR calculations. Compounds (+)-2 and (-)-2 exhibited remarkable antioxidative activities.

Isolation and Characterization of a Glycosyl Hydrolase Family 16 ß-Agarase from a Mangrove Soil Metagenomic Library.

A mangrove soil metagenomic library was constructed and a ß-agarase gene designated as AgaML was isolated by functional screening. The gene encoded for a 659-amino-acids polypeptide with an estimated molecular mass of 71.6 kDa. The deduced polypeptide sequences of AgaML showed the highest identity of 73% with the glycoside hydrolase family 16 ß-agarase from Microbulbifer agarilyticus in the GenBank database. AgaML was cloned and highly expressed in Escherichia coli BL21(DE3). The purified recombinant protein, AgaML, showed optimal activity at 50 °C and pH 7.0. The kinetic parameters of Km and Vmax values toward agarose were 4.6 mg·mL(-1) and 967.5 µM·min(-1)·mg(-1), respectively. AgaML hydrolyzed the ß-1,4-glycosidic linkages of agar to generate neoagarotetraose (NA4) and neoagarohexaose (NA6) as the main products. These characteristics suggest that AgaML has potential application in cosmetic, pharmaceuticals and food industries.

Alcohol dehydrogenases from Kluyveromyces marxianus: heterologous expression in Escherichia coli and biochemical characterization.

BACKGROUND: Kluyveromyces marxianus has recently become a species of interest for ethanol production since it can produce ethanol at high temperature and on a wide variety of substrates. However, the reason why this yeast can produce ethanol at high temperature is largely unknown. RESULTS: The ethanol fermentation capability of K. marxianus GX-UN120 at 40°Ð¡ was found to be the same as that of Saccharomyces cerevisiae at 34°Ð¡. Zymogram analysis showed that alcohol dehydrogenase 1 (KmAdh1) was largely induced during ethanol production, KmAdh4 was constitutively expressed at a lower level and KmAdh2 and KmAdh3 were almost undetectable. The genes encoding the four alcohol dehydrogenases (ADHs) were cloned from strain GX-UN120. Each KmADH was expressed in Escherichia coli and each recombinant protein was digested with enterokinase to remove the fusion protein. The optimum pH of the purified recombinant KmAdh1 was 8.0 and that of KmAdh2, KmAdh3 and KmAdh4 was 7.0. The optimum temperatures of KmAdh1, KmAdh2, KmAdh3 and KmAdh4 were 50, 45, 55 and 45°C, respectively. The K(m) values of the recombinant KmAdh1 and KmAdh2 were 4.0 and 1.2 mM for acetaldehyde and 39.7 and 49.5 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh1 and KmAdh2 were 114.9 and 21.6 µmol min⁻¹ mg⁻¹ for acetaldehyde and 57.5 and 1.8 µmol min⁻¹ mg⁻¹ for ethanol, respectively. KmAdh3 and KmAdh4 catalyze the oxidation reaction of ethanol to acetaldehyde but not the reduction reaction of acetaldehyde to ethanol, and the K(m) values of the recombinant KmAdh3 and KmAdh4 were 26.0 and 17.0 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh3 and KmAdh4 were 12.8 and 56.2 µmol min⁻¹ mg⁻¹ for ethanol, respectively. CONCLUSION: These data in this study collectively indicate that KmAdh1 is the primary ADH responsible for the production of ethanol from the reduction of acetaldehyde in K. marxianus. The relatively high optimum temperature of KmAdh1 may partially explain the ability of K. marxianus to produce ethanol at high temperature. Understanding the biochemical characteristics of KmAdhs will enhance our fundamental knowledge of the metabolism of ethanol fermentation in K. marxianus.

Cloning and characterization of a novel GH44 family endoglucanase from mangrove soil metagenomic library.

A novel endoglucanase gene, mgcel44, was isolated from a mangrove soil metagenomic library by functional-based screening. It encodes a 648-aa peptide with a catalytic domain of glycosyl hydrolase family 44. The deduced amino acid sequence of mgcel44 shares less than 50 % identity with endoglucanases in GenBank database. mgcel44 was cloned and overexpressed in Escherichia coli. The recombinant enzyme, MgCel44, has a molecular mass of 70.8 kDa as determined by SDS-PAGE. Its optimal pH and temperature for activity were 6 and 45 °C, respectively. It was highly active at 25-45 °C and pH 5-8. Its activity was enhanced in 0.5 M NaCl by >1.6-fold and stable up to 1.5 M NaCl. MgCel44 was resistant to several organic solvents and had high activity at 15 % (v/v) solvent after incubating for 24 h at 25 °C.

Biological soil crust elicits microbial community and extracellular polymeric substances restructuring to reduce the soil erosion on tropical island, South China Sea.

Soil erosion stands as the preeminent environmental concern globally, attaining heightened significance, particularly within islands where land resources prove notably scarce. Biological soil crusts, referred to as biocrusts, assume a pivotal ecological role in soil conservation. Notably, they augment the horizontal stability of the substrate through the exudation of microbial extracellular polymeric substances (EPS), thereby shielding the soil against shear stress, exemplified in the form of water erosion. While extant research has delved into the anti-erosion mechanisms of biocrusts in arid landscapes, a conspicuous lacuna persists in the exploration of coral island environments. In this study, we collected and assessed 30 samples encompassing dark biocrusts, light biocrusts, and bare soil to scrutinize the potential anti-erosion efficacy of tropical coral island biocrusts within the South China Sea. Employing a cohesive strength meter, we quantified soil shear stress across various stages of biocrust development, revealing a discernible enhancement in soil erosion resistance during the formation of biocrusts. Relative to the exposed bare soil, the soil shear stress exhibited an escalation from 0.33 N m-2 to 0.61 N m-2 and 1.31 N m-2 in the light biocrusts and dark biocrusts, respectively. Mechanistically, we assayed microbial EPS contents, exposing a positive correlation between EPS and soil anti-erodibility, encompassing extracellular protein and polysaccharide. Concurrently, bacterial abundance displayed a significant augmentation commensurate with biocrust formation and development. In pursuit of elucidating the origin of EPS, high-throughput amplicon sequencing was executed to identify microorganisms contributing to biocrust development. Correlation analysis discerned Cyanobacteria, Chloroflexi, Deinococcota, and Patescibacteria as potential microbials fostering EPS production and fortifying erosion resistance. Collectively, our study presents the first evidence that biocrust from tropical coral reef island in the South China Sea promotes resistance to soil erosion, pinpointing key EPS-producing microbials against soil erosion. The findings would provide insights for island environment restoration.

Dissemination of antibiotic resistance genes from the Pearl River Estuary to adjacent coastal areas.

The spread of antibiotic resistance genes (ARGs) is a growing concern over the world's various environments. Coastal environments may receive pollutants from land runoffs via estuaries. However, the impact of ARG contamination from estuarine regions to coastal areas is rarely reported. This study used high-throughput quantitative PCR to examine the diversity and abundance of ARGs in Pearl River Estuary (PRE) and adjacent coastal areas. We found that the distribution of ARGs in seawater exhibited the distance-decay phenomenon from the estuary to coastal areas, while the sediment samples did not exhibit an obvious distribution pattern. The estuarine water was found to be the hotspot of ARGs, with 74 ARG species detected and absolute abundance being 5.93 × 105 copies per mL, on average, while less species and lower abundance of ARGs were detected in coastal waters. Ordination analysis showed that estuarine ARG communities were significantly different from coastal ARG communities for water samples. SourceTracker analysis revealed that ARGs from the estuarine environment contributed only a minor fraction of ARG contamination to downstream coastal areas (1.5%-7.4% for water samples, and 0.7-1.8% for sediment samples), indicating the strong dilution effect of seawater. Mantel tests, redundancy analysis and random forest model analysis identified salinity, nutrients, microbial community structure and mobile genetic elements (MGEs) as important factors influencing ARG distribution. Partial least squares-path model revealed that, among all environmental factors, MGEs directly affected the distribution of ARGs, while other factors indirectly contributed by affecting the MGEs assemblage. Our study provides insight into the dissemination of ARGs from the PRE to adjacent coastal areas.

Radial Oxygen Loss from the Roots of Mangrove Seedlings Enhances the Removal of Polycyclic Aromatic Hydrocarbons.

The presence of polycyclic aromatic hydrocarbons (PAHs) in soil poses a significant global environmental concern, particularly in coastal wetlands. Mangrove ecosystems exhibit enormous potential in environmental purification; however, the underlying mechanisms involved in the degradation of pollutants (e.g., PAHs) remain ambiguous. In the present investigation, a soil pot experiment was conducted with the addition of pyrene to evaluate the effect of radial oxygen loss (ROL) from roots on PAH degradation using three mangrove seedlings (Rhizophora stylosa, Aegiceras corniculatum, and Avicennia marina). The results showed that mangrove plantation can significantly promote the efficiency of pyrene removal. As for the three mangrove species studied, the greatest removal rate (90.75%) was observed in the soils associated with A. marina, followed by A. corniculatum (83.83%) and R. stylosa (77.15%). The higher PAH removal efficiency of A. marina can be partially attributed to its distinctive root anatomical structure, characterized by a thin exodermis and high porosity, which facilitates ROL from the roots. The results from qPCR further demonstrate that ROL is beneficial for promoting the abundance of PAH-ring hydroxylating dioxygenase gene, leading to a higher removal efficiency. Additionally, Rhizobiales, Defferrisomatales, and Ardenticatenales may also play important roles in the process of pyrene degradation. In summary, this study provides evidence for elucidating the mechanism of PAH removal from the perspective of ROL, thereby contributing valuable insights for species selection during mangrove restoration and remediation.

Biocrust reduces the soil erodibility of coral calcareous sand by regulating microbial community and extracellular polymeric substances on tropical coral island, South China Sea.

Tropical coral islands assume a pivotal role in the conservation of oceanic ecosystem biodiversity. However, their distinctive environmental attributes and limited vegetation render them highly susceptible to soil erosion. The biological soil crust (biocrust), owing to its significant ecological role in soil stabilization and erosion prevention, is deemed an effective means of mitigating soil erosion on coral island. However, existing research on the mechanisms through which biocrusts resist soil erosion has predominantly concentrated on arid and semi-arid regions. Consequently, this study will specifically delve into elucidating the erosion-resistant mechanisms of biocrusts in tropical coral island environments, South China Sea. Specifically, we collected 16 samples of biocrusts and bare soil from Meiji Island. High-throughput amplicon sequencing was executed to analyze the microbial community, including bacteria, fungi, and archaea. Additionally, quantitative PCR was utilized to assess the abundance of the bacterial 16S rRNA, fungal ITS, archaeal 16S rRNA, and cyanobacterial 16S rRNA genes within these samples. Physicochemical measurements and assessments of extracellular polymeric substances (EPSs) were conducted to characterize the soil properties. The study reported a significantly decreased soil erodibility factor after biocrust formation. Compared to bare soil, soil erodibility factor decreased from 0.280 to 0.190 t h MJ-1 mm-1 in the biocrusts. Mechanistically, we measured the microbial EPS contents and revealed a negative correlation between EPS and soil erodibility factor. Consistent with increased EPS, the abundance of bacteria, fungi, archaea, and cyanobacteria were also detected significantly increased with biocrust formation. Correlation analysis detected Cyanobacteria, Chloroflexi, Deinococcota, and Crenarchaeota as potential microbials promoting EPSs and reducing soil erosion. Together, our study presents the evidence that biocrust from tropical coral island in the South China Sea promotes resistance to soil erosion, pinpointing key EPSs-producing microbials against soil erosion. The findings would provide insights for island soil restoration.

Bioaccumulation of emerging persistent organic pollutants in the deep-sea cold seep ecosystems: Evidence from chlorinated paraffin.

Persistent organic pollutants (POPs) are highly toxic and can accumulate in marine organisms, causing nonnegligible harm to the global marine ecosystem. The Cold seep is an essential marine ecosystem with the critical ecological function of maintaining the deep-sea carbon cycle and buffering global climate change. However, the environmental impact of emerging POPs in the deep-sea cold seep ecosystem is unknown. Here, we investigated the potential pollution of chlorinated paraffins (CPs) and their bioaccumulation in the cold seep ecosystem. High concentrations of CPs were detected in the cold seep ecosystems, where CPs bioaccumulated by the keystone species of deep-sea mussels can be released into the surface sediment and vertically migrate into the deeper sediment. Furthermore, more toxic CPs were accumulated from transforming other CPs in the cold seep ecosystem. Our study provides the first evidence that high concentrations of POPs are bioaccumulated by deep-sea mussels in the cold seep ecosystem, causing adverse ecological effects. The discovery of CPs bioaccumulation in the deep-sea cold seep ecosystem is a crucial mechanism affecting deep-sea carbon transport and cycling. This study has important guiding significance for revealing the deep-sea carbon cycle process, addressing global climate change, and making deep-sea ecological and environmental protection policies.

Role of extracellular polymeric substances in metal sequestration during mangrove restoration.

Extracellular polymeric substances (EPS) are widely observed in aquatic ecosystems, however the potential function of EPS on metal sequestration in mangrove wetlands is unclear. Thus, an ecological restoration area (including Sonneratia apetala, Kandelia obovata and unvegetated mudflat) was employed to assess the effect of mangrove reforestation on metal sequestration and the underlying roles played by EPS. The results showed that mangrove restoration directly promoted metal accumulation (e.g., Cr, Cu, Ni, Pb, and Zn) in sediments. However, alleviated metal bioavailability was detected after mangrove reforestation. The changes in metal accumulation and bioavailability were highly correlated with EPS and microbial composition. Mangrove restoration (especially for K. obovata reforestation) also significantly promoted EPS production, in which multiple metal-chelating functional groups (e.g., hydroxyl, carboxyl, and imino) were identified by Fourier infrared spectra. Moreover, the contents of EPS were positively correlated with metal accumulation but negatively correlated with metal bioavailability. The present data further illustrated that the enhancements of Gammaproteobacteria, Bacteroidia, Desulfobulbia, and Desulfobacteria might be important for EPS production. In summary, this is the first study to reveal that the presence of artificial mangroves might act as an efficient barrier in metal sequestration and immobilization by enhancing inherent microbial EPS.

Degradation potential of alkanes by diverse oil-degrading bacteria from deep-sea sediments of Haima cold seep areas, South China Sea.

Marine oil spills are a significant concern worldwide, destroying the ecological environment and threatening the survival of marine life. Various oil-degrading bacteria have been widely reported in marine environments in response to marine oil pollution. However, little information is known about culturable oil-degrading bacteria in cold seep of the deep-sea environments, which are rich in hydrocarbons. This study enriched five oil-degrading consortia from sediments collected from the Haima cold seep areas of the South China Sea. Parvibaculum, Erythrobacter, Acinetobacter, Alcanivorax, Pseudomonas, Marinobacter, Halomonas, and Idiomarina were the dominant genera. Further results of bacterial growth and degradation ability tests indicated seven efficient alkane-degrading bacteria belonging to Acinetobacter, Alcanivorax, Kangiella, Limimaricola, Marinobacter, Flavobacterium, and Paracoccus, whose degradation rates were higher in crude oil (70.3-78.0%) than that in diesel oil (62.7-66.3%). From the view of carbon chain length, alkane degradation rates were medium chains > long chains > short chains. In addition, Kangiella aquimarina F7, Acinetobacter venetianus F1, Limimaricola variabilis F8, Marinobacter nauticus J5, Flavobacterium sediminis N3, and Paracoccus sediminilitoris N6 were first identified as oil-degrading bacteria from deep-sea environments. This study will provide insight into the bacterial community structures and oil-degrading bacterial diversity in the Haima cold seep areas, South China Sea, and offer bacterial resources to oil bioremediation applications.

Mangrove restoration promotes the anti-scouribility of the sediments by modifying inherent microbial community and extracellular polymeric substance.

Coastal erosion will aggravate the loss of shorelines and threaten the safety of coastal engineering facilities. Mangrove is often considered as an efficient coastal guard; however the mechanisms involved in anti-scouribility ascribed to mangrove are still poorly understood. Thus, two artificial mangrove forests (including exotic Sonneratia apetala and native Kandelia obovata) and an unvegetated mudflat control were selected to explore the potential function of microbial extracellular polymeric substance (EPS) on the anti-scouribility of the sediments. A cohesive strength meter was used for the analysis of anti-scouribility, while a sequential extraction and 16S high-throughput sequencing were employed to evaluate the changes in EPS and microbial community driven by mangrove restoration. Principal component, redundancy, and two-matrix correlation heatmap analyses were performed for the analyses of the correlations among shear stress, EPS, microbes, and soil properties. The results showed an obvious enhancement of anti-scouribility after mangrove restoration. Compared to those of unvegetated mudflat, shear stress increased from 1.94 N/m2 to 3.26 and 4.93 N/m2 in the sediments of S. apetala and K. obovata stands, respectively. Mangrove restoration also promoted EPS content in the sediments, irrespective of EPS components and sub-fractions. Both extracellular protein and polysaccharide were found to be positively correlated with anti-scouribility. Coinciding with increased anti-scouribility and EPS, increased bacterial abundances were also detected in the sediments after mangrove restoration (especially K. obovata), whereas Proteobacteria and Bacteroides may be important and influential for EPS secretion and anti-scouribility promotion. Nevertheless, increased total organic carbon, total nitrogen and total phosphorus induced by mangrove restoration may also partially contribute to improvement of anti-scouribility. In conclusion, this is the first study to provide evidence for a link between mangrove restoration and increased EPS which improve resistance to scouring. The present study provides a novel perspective on the revealing of the function of mangrove on erosion mitigation.

Characteristics of Microbial Community and Function With the Succession of Mangroves.

In this study, 16S high-throughput and metagenomic sequencing analyses were employed to explore the changes in microbial community and function with the succession of mangroves (Sonneratia alba, Rhizophora apiculata, and Bruguiera parviflora) along the Merbok river estuary in Malaysia. The sediments of the three mangroves harbored their own unique dominant microbial taxa, whereas R. apiculata exhibited the highest microbial diversity. In general, Gammaproteobacteria, Actinobacteria, Alphaproteobacteria, Deltaproteobacteria, and Anaerolineae were the dominant microbial classes, but their abundances varied significantly among the three mangroves. Principal coordinates and redundancy analyses revealed that the specificity of the microbial community was highly correlated with mangrove populations and environmental factors. The results further showed that R. apiculata exhibited the highest carbon-related metabolism, coinciding with the highest organic carbon and microbial diversity. In addition, specific microbial taxa, such as Desulfobacterales and Rhizobiales, contributed the highest functional activities related to carbon metabolism, prokaryote carbon fixation, and methane metabolism. The present results provide a comprehensive understanding of the adaptations and functions of microbes in relation to environmental transition and mangrove succession in intertidal regions. High microbial diversity and carbon metabolism in R. apiculata might in turn facilitate and maintain the formation of climax mangroves in the middle region of the Merbok river estuary.

Asperorydines N-P, three new cyclopiazonic acid alkaloids from the marine-derived fungus Aspergillus flavus SCSIO F025.

Three new tricyclic cyclopiazonic acid (CPA) related alkaloids asperorydines N-P (1-3), together with six known compounds (4-9) were isolated and characterized from the fungus Aspergillus flavus SCSIO F025 derived from the deep-sea sediments of South China Sea. The structures including absolute configurations of 1-3 were deduced from spectroscopic data, X-ray diffraction analysis, and electronic circular dichroism (ECD). All compounds were evaluated for the antioxidative activities against DPPH, cytotoxic activities against four tumor cell lines (SF-268, HepG-2, MCF-7, and A549), and antimicrobial activities. Compound 9 showed significant radical scavenging activities against DPPH with an IC50 value of 62.23 µM and broad-spectrum cytotoxicities against four tumor cell lines with IC50 values ranging from 24.38 to 48.28 µM. Furthermore, compounds 4-9 exhibited weak antimicrobial activities against E scherichia coli, and compound 9 also showed antibacterial activity against Bacillus thuringiensis, Micrococcus lutea, Staphylococcus aureus, Bacillus subtilis, Methicillin resistant Staphylococcus aureus.

Community differentiation of bacterioplankton in the epipelagic layer in the South China Sea.

The South China Sea (SCS) is the largest marginal sea in the western tropical Pacific Ocean and is characterized by complex physicochemical environments. To date, the biogeographic patterns of the microbial communities have rarely been reported at a basin scale in the SCS. In this study, the bacterial assemblages inhabiting the epipelagic zone across 110°E to 119°E along 14°N latitude were uncovered. The vertical stratification of both bacterial taxa and their potential functions were revealed. These results suggest that the water depth-specific environment is a driver of the vertical bacterioplankton distribution. Moreover, the bacterial communities were different between the eastern stations and the western stations, where the environmental conditions were distinct. However, the mesoscale eddy did not show an obvious effect on the bacterial community due to the large distance between the sampling site and the center of the eddy. In addition to the water depth and longitudinal location of the samples, the heterogeneity of the phosphate and salinity concentrations also significantly contributed to the variance in the epipelagic bacterial community in the SCS. To the best of our knowledge, this study is the first to report that the variability in epipelagic bacterioplankton is driven by the physicochemical environment at the basin scale in the SCS. Our results emphasize that the ecological significance of bacterioplankton can be better understood by considering the relationship between the biogeographic distribution of bacteria and the oceanic dynamics processes.

Cloning, Expression, and Characterization of a Cold-Active and Organic Solvent-Tolerant Lipase from Aeromicrobium sp. SCSIO 25071.

The gene encoding lipase (Lip98) from Aeromicrobium sp. SCSIO 25071 was cloned and functionally expressed in Escherichia coli. Lip98 amino acid sequence shares the highest (49%) identity to Rhodococcus jostii RHA1 lipase and contains a novel motif (GHSEG), which is different from other clusters in the lipase superfamily. The recombinant lipase was purified to homogeneity with Ni-NTA affinity chromatography. Lip98 showed an apparent molecular mass of 30 kDa on SDS gel. The optimal temperature and pH value for enzymatic activity were recorded at 30°C and 7.5, respectively. Lip98 exhibited high activity at low temperatures with 35% maximum activity at 0°C and good stability at temperatures below 35°C. Its calculated activation energy was 4.12 kcal/mol at the low temperature range of 15-30°C. Its activity was slightly affected by some metal ions such as K(+), Ca(2+), and Na(+). The activity of Lip98 was increased by various organic solvents such as DMSO, ethanol, acetone, and hexane with the concentration of 30% (v/v) and retained more than 30% residual activity in neat organic solvent. The unique characteristics of Lip98 imply that it is a promising candidate for industrial application as a nonaqueous biocatalyst and food additive.

Purification, characterization, and gene cloning of a cold-adapted thermolysin-like protease from Halobacillus sp. SCSIO 20089.

Marine sediment is a distinctive habitat of cold enzyme producing bacteria. A protease producing strain Halobacillus sp. SCSIO 20089 was isolated from a marine sediment of South China Sea. Using chromatographic techniques, the extracellular protease was purified to homogeneity from the culture supernatant. The purified protease exhibited maximal activity at 30°C, pH 8.0, and remained more than 20% of its activity at 0°C. Its activation energy was calculated to be 34.4 kJ/mol, suggesting it is a cold-adapted protease. Based on the N-terminal amino acid sequence of the purified enzyme, full gene encoding the enzyme was obtained by combination of degenerate primer PCR and hiTAIL-PCR. The deduced amino acid sequence showed 57% and 52% identity with mesothermal and thermophilic protease in thermolysin family respectively. All these indicate the enzyme is a unique cold-active thermolysin-like protease with potential in both basic research and industrial application areas.