Bacterial Extracellular Enzymes Research Articles

Resin Acid Copper Salt, an Interesting Chemical Pesticide, Controls Rice Bacterial Leaf Blight by Regulating Bacterial Biofilm, Motility, and Extracellular Enzymes.

Bacterial virulence plays an important role in infection. Antibacterial virulence factors are effective for preventing crop bacterial diseases. Resin acid copper salt as an effective inhibitor exhibited excellent anti-Xanthomonas oryzae pv. oryzae (Xoo) activity with an EC50 of 50.0 μg mL-1. Resin acid copper salt (RACS) can reduce extracellular polysaccharides' (EPS's) biosynthesis by down-regulating gumB relative expression. RACS can also effectively inhibit the bio-mass of Xoo biofilm. It can reduce the activity of Xoo extracellular amylase at a concentration of 100 μg mL-1. Meanwhile, the results of virtual computing suggested that RACS is an enzyme inhibitor. RACS displayed good curative activity with a control effect of 38.5%. Furthermore, the result of the phytotoxicity assessment revealed that RACS exhibited slight toxicity compared with the control at a concentration of 200 μg mL-1. The curative effect was increased to 45.0% using an additional antimicrobial agent like orange peel essential oil. RACS markedly inhibited bacterial pathogenicity at a concentration of 100 μg mL-1 in vivo.

Unprecedented Diversity of the Glycoside Hydrolase Family 70: A Comprehensive Analysis of Sequence, Structure, and Function.

The glycoside hydrolase family 70 (GH70) contains bacterial extracellular multidomain enzymes, synthesizing α-glucans from sucrose or starch-like substrates. A few dozen have been biochemically characterized, while crystal structures cover only the core domains and lack significant parts of auxiliary domains. Here we present a systematic overview of GH70 enzymes and their 3D structural organization and bacterial origin. A representative set of 234 permuted and 25 nonpermuted GH70 enzymes was generated, covering 12 bacterial families and 3 phyla and containing 185 predicted glucansucrases (GS), 15 branching sucrases (BrS), 8 "twin" GS-BrSs, and 51 α-glucanotransferases (α-GT). Analysis of AlphaFold models of all 259 entries showed that, apart from the core domains, the structural variation regarding auxiliary domains is far greater than anticipated, with nine different domain types. We analyzed the phylogenetic distribution and discuss the possible roles of auxiliary domains as well as possible correlations between enzyme specificity, auxiliary domain type, and bacterial origin.

Enrichment, domestication, degradation, adaptive mechanism, and nicosulfuron bioremediation of bacteria consortium YM21

Nicosulfuron (NSR), a sulfonylurea herbicide, can easily enter water bodies, affecting aquatic life and human safety. Here, bacteria consortium YM2 was enriched and domesticated from the active sludge of a pesticide plant for the bioremediation of NSR wastewater. The response surface methodology data revealed that under optimal incubation conditions: 9.41 g L–1 maltodextrin, 21.37 g L–1 yeast extract, and 12.45 g L–1 NaCl, YM2 bacteria consortium degraded 97.49% of NSR within 4 d. The optimal degradation conditions were temperature 30°C, pH 6.0, inoculum 1%, and initial NSR concentration 20 mg L–1), The degradation system was tolerant to heavy metal ions such as Cd2+, Pb2+, Ni2+, and Zn2+ and mainly occurred through the bacterial extracellular enzymes (92.17%). Mechanistically, during the degradation process, reactive oxygen species, oxidative stress, cell membrane permeability, cell surface hydrophobicity, and apoptosis rate first increased and then decreased. Also, the expression of biofilm formation-related genes luxS, waaE, spo0A, and wza varied with time and concentration. NSR wastewater and soil were degraded to 1.92 mg L–1 and 2.72 mg L–1, respectively. In a simulated wastewater treatment unit (hydraulic retention time 12 h), YM2 degraded 84.55% of NSR after 10 d. This study provides a theoretical basis for the microbial remediation of NSR.

Characteristics of bacterial community and extracellular enzymes in response to atrazine application in black soil

The ecological functioning of black soil largely depends on the activities of various groups of microorganisms. However, little is known about how atrazine, a widely used herbicide with known harmful effects on the environment, influences the microbial ecology of black soil, and the extracellular enzymes related to the carbon, nitrogen and phosphorus cycles. Here, we evaluated the change in extracellular enzymes and bacterial community characteristics in black soil after exposure to various concentrations of atrazine. Low concentrations of applied atrazine (10 – 20 mg kg−1) were almost completely degraded after 120 days. At high concentrations (80 – 100 mg kg−1), about 95% of the applied atrazine was degraded over the same period. Additionally, linear fitting of data indicated that the total enzymatic activity index (TEI) and bacterial α-diversity index were negatively correlated with atrazine applied concentration. The atrazine had a greater effect on bacterial beta diversity after 120 days, which differentiated species clusters treated with low and high atrazine concentrations. Soil bacterial community structure and function were affected by atrazine, especially at high atrazine concentrations (80 – 100 mg kg−1). Key microorganisms such as Sphingomonas and Nocardioides were identified as biomarkers for atrazine dissipation. Functional prediction indicated that most metabolic pathways might be involved in atrazine dissipation. Overall, the findings enhance our understanding of the factors driving atrazine degradation in black soil and supports the use of biomarkers as indicators of atrazine dissipation.

Exoenzyme Profiling of Soil Bacteria from Thattekad Bird Sanctuary for Bioprospection

Thattekad bird sanctuary, located in the Western Ghats of Kerala, India, which hosts an unexplored microbial community, is selected for the present investigation. Microbes play a major role in mineral recycling and nutrient absorption by the flora and fauna in the habitat. Various bacterial extracellular enzymes facilitate all these activities. The increasing demand for microbial enzymes in favor of green technology encouraged us to focus on exoenzyme profiling of bacterial isolates from forest soil samples. The present study is aimed at the screening and identification of exoenzyme producing soil bacterial strains isolated from evergreen forests and moist deciduous forests of Thattekad bird sanctuary. In this study, only multienzyme producing bacteria were selected for detailed analysis because such bacteria are highly relevant in multi-enzyme dependent processes such as biowaste degradation. We screened for nine hydrolytic exoenzymes namely, amylase, cellulase, ligninase, pectinase, xylanase, caseinase, gelatinase, esterase and lipase, and identified 79 multienzyme-producing bacterial strains, mostly belonging to phylum Firmicutes and Proteobacteria. Firmicutes from evergreen forests and moist deciduous forests produced a greater number of enzymes compared to Proteobacteria. Also, bacterial strains isolated from evergreen forest soil produced more enzymes compared to moist deciduous forest. Bacillus amyloliquefaciens strain TBS040 isolated from moist deciduous forest soil was found to produce all the nine enzymes screened. Enzymatic hydrolysis of biowaste using cell free crude enzyme extract from Bacillus velezensis strain TBS064 resulted in enhanced bioethanol production. These findings highlight the importance of screening unexplored habitats for the identification of novel strains, which can contribute to the future of green technology.

A resveratrol oligomer, hopeaphenol suppresses virulence activity of Pectobacterium atrosepticum via the modulation of the master regulator, FlhDC.

Pectobacterium atrosepticum (P. atrosepticum: Pba) which causes potato soft rot and blackleg is a notorious plant pathogen worldwide. Discovery of new types of antimicrobial chemicals that target specifically to virulence factors such as bacterial motility and extracellular enzymes is required for protecting crops from pathogenic infection. A transcriptomic analysis of Pba upon hopeaphenol treatment revealed that bacterial motility-related gene expression, including a master regulator flhDC genes, was significantly influenced by hopeaphenol. We further generated a double knock-out mutant of flhDC genes by CRISPR/Cas9 system and confirmed phenotypic changes in bacterial motility, transcription of extracellular enzymes, and disease development consistent with the result of wild-type treated with hopeaphenol. The hopeaphenol-treated Pba strains, wild-type, double mutant, and complemented strain were unable to secrete the enzymes in vitro, while ΔflhDC double mutant strain reduced the secretion. Thus, our study supports that FlhDC is essential for the virulence of Pba, and proposes that hopeaphenol modulates FlhDC-dependent virulence pathways, suggesting a potential of hopeaphenol as an anti-virulence agent to manage potato soft rot and blackleg diseases.

Exploring an Innovative Strategy for Suppressing Bacterial Plant Disease: Excavated Novel Isopropanolamine-Tailored Pterostilbene Derivatives as Potential Antibiofilm Agents.

Bacterial biofilms are the root cause of persistent and chronic phytopathogenic bacterial infections. Therefore, developing novel agrochemicals that target the biofilm of phytopathogenic bacteria has been regarded as an innovative tactic to suppress their invasive infection or decrease bacterial drug resistance. In this study, a series of natural pterostilbene (PTE) derivatives were designed, and their antibacterial potency and antibiofilm ability were assessed. Notably, compound C1 displayed excellent antibacterial potency in vitro, affording an EC50 value of 0.88 μg mL-1 against Xoo (Xanthomonas oryzae pv. oryzae). C1 could significantly reduce biofilm formation and extracellular polysaccharides (EPS). Furthermore, C1 also possessed remarkable inhibitory activity against bacterial extracellular enzymes, pathogenicity, and other virulence factors. Subsequently, pathogenicity experiments were further conducted to verify the above primary outcomes. More importantly, C1 with pesticide additives displayed excellent control efficiency. Given these promising profiles, these pterostilbene derivatives can serve as novel antibiofilm agents to suppress plant pathogenic bacteria.

Experimental evidence for long-term coexistence of copiotrophic and oligotrophic bacteria in pelagic surface seawater.

Most marine copiotrophic bacteria can produce extracellular enzymes to degrade biopolymers into bio-available smaller solutes, while oligotrophic bacteria usually cannot. Bacterial extracellular enzymes and enzymatic products can be a common resource that could be utilized by both copiotrophs and oligotrophs; when present, oligotrophs may outcompete the enzyme-producing copiotrophs. However, copiotrophs and oligotrophs consistently coexist in the ocean. How they maintain coexistence has still not been experimentally studied. In this study, the interaction and coexistence of a copiotroph and an oligotroph, isolated from the same surface seawater sample and utilizing the same proteinaceous substrate, were experimentally investigated. The copiotroph could secrete extracellular proteases to degrade and then utilize the proteinaceous substrate. The oligotroph was unable to utilize the proteinaceous substrate by itself, but could grow by using the hydrolysate amino acids. The copiotroph outcompeted the oligotroph by adsorbing the amino acids quickly and having a higher growth rate in the rich medium. The oligotroph survived by adapting to low concentration of nutrients. The copiotroph and oligotroph were able to maintain long-term (up to 142 days) coexistence in the laboratory. This study indicates that differences in the utilization of different concentrations of nutrients can drive the coexistence of marine copiotrophs and oligotrophs.

Binding of cloned Cel enzymes on clay minerals related to the pI of the enzymes and database survey of cellulases of soil bacteria for pI.

The Cel genes from Bacillus licheniformis MSB03 were cloned and expressed to investigate binding ability on clay minerals and sea sand at pH ranging 3 to 9. FTIR analysis has been done to characterize bound enzymes on clay minerals. Subsequent, surveying of NCBI database for extracellular enzymes of soil bacteria was carried out. Among the five cloned Cel enzymes assayed for binding to clay minerals, only Cel5H enzyme had the binding ability. Enzyme Cel5H exhibited highest binding to montmorillonite followed by kaolinite and sea sand. Interestingly, Cel5H had higher pI value of 9.24 than other proteins (5.2-5.7). Cel5H binding to montmorillonite was shown to be negatively affected below pH 3 and above pH 9. Infrared absorption spectra of the Cel5H-montmorillonite complexes showed distinct peaks for clay minerals and bound proteins. Furthermore, database survey of soil bacterial extracellular enzymes revealed that Bacillus species enzymes had higher pI than other soil bacterial enzymes.

Short-term warming shifts microbial nutrient limitation without changing the bacterial community structure in an alpine timberline of the eastern Tibetan Plateau

The forest-alpine ecotone is consideredsusceptible to climate change. Addressing the effects of climate warming on the structure and function of the soil microbial community will provide insights into carbon (C) and nutrient cycling under climate change. A short-term open-top chamber (OTC) experiment was conducted in the alpine timberline of the eastern Tibetan Plateau to test the effects of experimental warming on the soil bacterial community composition, extracellular enzymes, ecoenzymatic stoichiometry and soil physicochemical properties. The results showed that there were no significant changes in the soil bacterial alpha-diversity, composition or community structure in response to soil warming. However, during the early growing season, two carbon-degrading enzymes (β-glucosidase, BG; peroxidase, POD) and microbial carbon limitation decreased with soil warming in the mineral soil layer, which might be a result of an increasing accumulation of soil C. We also observed that C:P enzyme ratios decreased and the relative microbial P limitation increased with soil warming in the organic soil layer. Our db-RDA results demonstrate that both available nutrient pools (dissolved organic carbon and total dissolved nitrogen) and total nutrient concentrations (soil organic carbon and total nitrogen), in addition to soil pH and moisture content, affected soil bacterial communities. These findings suggest that short-term soil warming may change the microbial C and P demand, although bacteria showed temperature adaptability to soil warming.

Key extracellular enzymes triggered high-efficiency composting associated with bacterial community succession

A consortium of key bacterial taxa plays critical roles in the composting process. In order to elucidate the identity and mechanisms by which specific bacterial species drive high-efficiency composting, the succession of key bacterial consortia and extracellular enzymes produced during the composting process were monitored in composting piles with varying initial C/N ratios. Results showed that C/N ratios of 25 and 35 enhanced composting efficiency through elevated temperatures, higher germination indices, enhanced cellulose and hemicellulose degradation, and higher cellulase and dehydrogenase activities. The activities of cellulase and β-glucosidase, cellulase and protease, and cellulase and β-glucosidase exhibited significant relationships with bacterial community composition within the mesophilic, thermophilic, and mature phases, respectively. Putative key taxa, linked to a higher composting efficiency, such as Nonomuraea, Desemzia, Cellulosimicrobium, Virgibacillus, Clostridium, and Achromobacter, exhibited significantly positive relationships with extracellular enzyme activities, suggesting a significant contribution to these taxa to the development of composting maturity.

Extracellular Enzyme Activity Profile in a Chemically Enhanced Water Accommodated Fraction of Surrogate Oil: Toward Understanding Microbial Activities After the Deepwater Horizon Oil Spill.

Extracellular enzymes and extracellular polymeric substances (EPS) play a key role in overall microbial activity, growth and survival in the ocean. EPS, being amphiphilic in nature, can act as biological surfactant in an oil spill situation. Extracellular enzymes help microbes to digest and utilize fractions of organic matter, including EPS, which can stimulate growth and enhance microbial activity. These natural processes might have been altered during the 2010 Deepwater Horizon oil spill due to the presence of hydrocarbon and dispersant. This study aims to investigate the role of bacterial extracellular enzymes during exposure to hydrocarbons and dispersant. Mesocosm studies were conducted using a water accommodated fraction of oil mixed with the chemical dispersant, Corexit (CEWAF) in seawater collected from two different locations in the Gulf of Mexico and corresponding controls (no additions). Activities of five extracellular enzymes typically found in the EPS secreted by the microbial community – α- and β-glucosidase, lipase, alkaline phosphatase, leucine amino-peptidase – were measured using fluorogenic substrates in three different layers of the mesocosm tanks (surface, water column and bottom). Enhanced EPS production and extracellular enzyme activities were observed in the CEWAF treatment compared to the Control. Higher bacterial and micro-aggregate counts were also observed in the CEWAF treatment compared to Controls. Bacterial genera in the order Alteromonadaceae were the most abundant bacterial 16S rRNA amplicons recovered. Genomes of Alteromonadaceae commonly have alkaline phosphatase and leucine aminopeptidase, therefore they may contribute significantly to the measured enzyme activities. Only Alteromonadaceae and Pseudomonadaceae among bacteria detected here have higher percentage of genes for lipase. Piscirickettsiaceae was abundant; genomes from this order commonly have genes for leucine aminopeptidase. Overall, this study provides insights into the alteration to the microbial processes such as EPS and extracellular enzyme production, and to the microbial community, when exposed to the mixture of oil and dispersant.

Depth-Resolved Variations of Cultivable Bacteria and Their Extracellular Enzymes in the Water Column of the New Britain Trench.

Marine microorganisms and their extracellular enzymes (ECEs) play an important role in the remineralization of organic material by hydrolyzing high-molecular-weight substrates to sizes sufficiently small to be transported through cell membrane, yet the diversity of the enzyme-producing bacteria and the types of ECEs involved in the degradation process are largely unknown. In this work, we investigated the diversity of cultivable bacteria and their ECEs and the potential activities of aminopeptidase in the water column at eight different depths of the New Britain Trench. There was a great diversity of cultivable bacteria and ECEs, and depth appears an important driver of the diversity. The 16S rRNA sequence analysis revealed that the cultivable bacteria were affiliated mostly with the phyla Proteobacteria and Actinobacteria, and the predominant genera were Pseudoalteromonas (62.7%) and Halomonas (17.3%). Moreover, 70.7% of the isolates were found to produce hydrolytic zone on casein and gelatin plates, in which Pseudoalteromonas was the predominant group, exhibiting relatively high protease production. Inhibitor analysis showed that the extracellular proteases from the isolated bacteria were serine proteases in the surface water and metalloproteases in the deep water. Meanwhile, the Vmax and Km of aminopeptidase exhibited a maximum in the surface water and low values in the deep bathy- and abyssopelagic water, indicating lower rates of hydrolysis and higher substrate affinity in the deeper waters. These results shed new insights into the diversity of the cultivable bacteria and bacterial ECEs and their likely biogeochemical functions in the trench environment.

Nano-TiO2 affects Cu speciation, extracellular enzyme activity, and bacterial communities in sediments

In aquatic ecosystems, titanium dioxide nanoparticles (nano-TiO2) coexist with heavy metals and influence the existing forms and toxicities of the metal in water. However, limited information is available regarding the ecological risk of this coexistence in sediments. In this study, the effect of nano-TiO2 on Cu speciation in sediments was investigated using sequential extraction. The microcosm approach was also employed to analyze the effects of the coexistence of nano-TiO2 and Cu on extracellular enzyme activity and bacterial communities in sediments. Results showed that nano-TiO2 decreased the organic matter-bound fraction of Cu and increased the corresponding residual fraction Cu. As a result, speciation of exogenous Cu in sediments changed. During the course of the 30-day experiment, the presence of nano-TiO2 did not affect Cu-induced changes in bacterial community structure. However, the coexistence of nano-TiO2 and Cu restrained the activity of bacterial extracellular enzymes, such as alkaline phosphatase and β-glucosidase. The degree of inhibition also varied because of the different properties of extracellular enzymes. This research highlighted the importance of understanding and predicting the effects of the coexistence of nanomaterials and other pollutants in sediments.

Assessing approaches to determine the effect of ocean acidification on bacterial processes

Abstract. Bacterial extracellular enzymes play a significant role in the degradation of labile organic matter and nutrient availability in the open ocean. Although bacterial production and extracellular enzymes may be affected by ocean acidification, few studies to date have considered the methodology used to measure enzyme activity and bacterial processes. This study investigated the potential artefacts in determining the response of bacterial growth and extracellular glucosidase and aminopeptidase activity to ocean acidification as well as the relative effects of three different acidification techniques. Tests confirmed that the observed effect of pH on fluorescence of artificial fluorophores, and the influence of the MCA fluorescent substrate on seawater sample pH, were both overcome by the use of Tris buffer. In experiments testing different acidification methods, bubbling with CO2 gas mixtures resulted in higher β-glucosidase activity and 15–40 % higher bacterial abundance, relative to acidification via gas-permeable silicon tubing and acid addition (HCl). Bubbling may stimulate carbohydrate degradation and bacterial growth, leading to the incorrect interpretation of the impacts of ocean acidification on organic matter cycling.

Suspended marine particulate proteins in coastal and oligotrophic waters

Metaproteomic analyses were performed on suspended sediments collected in one coastal environment (Washington margin, Pacific Ocean, n=5) and two oligotrophic environments (Atlantic Ocean near BATS, n=5, and Pacific Ocean near HOTS, n=5). Using a database of 2.3 million marine proteins developed using the NCBI database, 443 unique peptides were detected from which 363 unique proteins were identified. Samples from the euphotic zone contained on average 2-3x more identifiable proteins than deeper waters (150-1500m) and these proteins were predominately from photosynthetic organisms. Diatom peptides dominate the spectra of the Washington margin while peptides from cyanobacteria, such as Synechococcus sp. dominated the spectra of both oligotrophic sites. Despite differences in the exact proteins identified at each location, there is good agreement for protein function and cellular location. Proteins in surface waters code for a variety of cellular functions including photosynthesis (24% of detected proteins), energy production (10%), membrane production (9%) and genetic coding and reading (9%), and are split 60-40 between membrane proteins and intracellular cytoplasmic proteins. Sargasso Sea surface waters contain a suite of peptides consistent with proteins involved in circadian rhythms that promote both C and N fixation at night. At depth in the Sargasso Sea, both muscle-derived myosin protein and the muscle-hydrolyzing proteases deseasin MCP-01 and metalloprotease Mcp02 from γ-proteobacteria were observed. Deeper waters contain peptides predominately sourced from γ-proteobacteria (37% of detected proteins) and α-proteobacteria (26%), although peptides from membrane and photosynthetic proteins attributable to phytoplankton were still observed (13%). Relative to surface values, detection frequencies for bacterial membrane proteins and extracellular enzymes rose from 9 to 16 and 2 to 4% respectively below the thermocline and the overall balance between membrane proteins and intracellular proteins grows to an approximate 75-25 split. Unlike the phytoplankton membrane proteins, which are detrital in nature, the bacterial protein suite at depth is consistent with living biomass.

Labile and Recalcitrant Organic Matter Utilization by River Biofilm Under Increasing Water Temperature

Microbial biofilms in rivers contribute to the decomposition of the available organic matter which typically shows changes in composition and bioavailability due to their origin, seasonality, and watershed characteristics. In the context of global warming, enhanced biofilm organic matter decomposition would be expected but this effect could be specific when either a labile or a recalcitrant organic matter source would be available. A laboratory experiment was performed to mimic the effect of the predicted increase in river water temperature (+4°C above an ambient temperature) on the microbial biofilm under differential organic matter sources. The biofilm microbial community responded to higher water temperature by increasing bacterial cell number, respiratory activity (electron transport system) and microbial extracellular enzymes (extracellular enzyme activity). At higher temperature, the phenol oxidase enzyme explained a large fraction of respiratory activity variation suggesting an enhanced microbial use of degradation products from humic substances. The decomposition of hemicellulose (β-xylosidase activity) seemed to be also favored by warmer conditions. However, at ambient temperature, the enzymes highly responsible for respiration activity variation were β-glucosidase and leu-aminopeptidase, suggesting an enhanced microbial use of polysaccharides and peptides degradation products. The addition of labile dissolved organic carbon (DOC; dipeptide plus cellobiose) caused a further augmentation of heterotrophic biomass and respiratory activity. The changes in the fluorescence index and the ratio Abs(250)/total DOC indicated that higher temperature accelerated the rates of DOC degradation. The experiment showed that the more bioavailable organic matter was rapidly cycled irrespective of higher temperature while degradation of recalcitrant substances was enhanced by warming. Thus, pulses of carbon at higher water temperature might have consequences for DOC processing.

Efficient E. coli expression systems for the production of recombinant β-mannanases and other bacterial extracellular enzymes

Two Escherichia coli expression systems based on T7 RNA polymerase promoter (pET system) and tac promoter (pFLAG system) have been used for the production and secretion of recombinant b-mannanases from Bacillus sp. Both E. coli OmpA signal peptide and native Bacillus signal peptide could be used efficiently for the secretion of recombinant enzymes into periplasmic space and culture media. The genes could be induced for over-expression with 0.1 - 1 mM isopropyl-β-D- 1-thiogalactopyranoside (IPTG) when the OD600 of the culture broth reached 0.6-1.5. The recombinant enzymes could be harvested from whole cell lysate, perimplasmic extract, or culture broth after induction for 4 - 20 hours. Since the enzyme is C-terminally tagged with hexahistidine, the recombinant enzymes could be conveniently purified to apparent homogeneity by one-step immobilized-metal affinity chromatography (IMAC) using Ni-NTA resins. The characteristics of purified recombinant b-mannanases from B. licheniformis and B. subtilis, which share 78% amino acid identity, are slightly different. These systems should be applicable for the production of various recombinant bacterial extracellular enzymes.

Acidification increases microbial polysaccharide degradation in the ocean

Abstract. With the accumulation of anthropogenic carbon dioxide (CO2), a proceeding decline in seawater pH has been induced that is referred to as ocean acidification. The ocean's capacity for CO2 storage is strongly affected by biological processes, whose feedback potential is difficult to evaluate. The main source of CO2 in the ocean is the decomposition and subsequent respiration of organic molecules by heterotrophic bacteria. However, very little is known about potential effects of ocean acidification on bacterial degradation activity. This study reveals that the degradation of polysaccharides, a major component of marine organic matter, by bacterial extracellular enzymes was significantly accelerated during experimental simulation of ocean acidification. Results were obtained from pH perturbation experiments, where rates of extracellular α- and β-glucosidase were measured and the loss of neutral and acidic sugars from phytoplankton-derived polysaccharides was determined. Our study suggests that a faster bacterial turnover of polysaccharides at lowered ocean pH has the potential to reduce carbon export and to enhance the respiratory CO2 production in the future ocean.

Inter-kingdom encounters: recent advances in molecular bacterium–fungus interactions

Interactions between bacteria and fungi are well known, but it is often underestimated how intimate and decisive such associations can be with respect to behaviour and survival of each participating organism. In this article we review recent advances in molecular bacterium-fungus interactions, combining the data of different model systems. Emphasis is given to the positive or negative consequences these interactions have on the microbe accommodating plants and animals. Intricate mechanisms of antagonism and tolerance have emerged, being as important for the biological control of plants against fungal diseases as for the human body against fungal infections. Bacterial growth promoters of fungal mycelium have been characterized, and these may as well assist plant-fungus mutualism as disease development in animals. Some of the toxins that have been previously associated with fungi are actually produced by endobacteria, and the mechanisms that lie behind the maintenance of such exquisite endosymbioses are fascinating. Bacteria do cause diseases in fungi, and a synergistic action between bacterial toxins and extracellular enzymes is the hallmark of such diseases. The molecular study of bacterium-fungus associations has expanded our view on microbial communication, and this promising field shows now great potentials in medicinal, agricultural and biotechnological applications.