Salaquimonas pukyongi gen. nov., sp. nov., a novel bacterium within the family Phyllobacteriaceae.

A novel aerobic, Gram-negative bacterial strain, RR3-28T, was isolated from a seawater recirculating aquaculture system in Busan, Republic of Korea. Cells were rod-shaped, non-motile, oxidase-positive, catalase-negative and grew optimally at 25-30 °C, pH 8.5 and 3 % (w/v) NaCl. Based on the results of phylogenetic analysis, strain RR3-28T was most closely related to Zhengella mangrovi X9-2-2T within the family Phyllobacteriaceae with 95.97 % 16S rRNA gene sequence similarity. The major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c, 71.1 %) and 11-methyl C18 : 1ω7c (14.4 %). The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine and unidentified aminolipids. The predominant quinone was ubiquinone Q-10 and the DNA G+C content was 58.6 mol%. On the basis of its phenotypic and genotypic characteristics, strain RR3-28T represents a novel genus and species belonging to the family Phyllobacteriaceae, for which the name Salaquimonas pukyongi gen. nov., sp. nov. is proposed. The type strain of the species is RR3-28T (=KCTC 52649T=DSM 107947T).

Industrial robustness linked to the gluconolactonase from Zymomonas mobilis.

The physiological characteristics and the potential gluconolactone production of the gluconolactonase-deficient strain, Zymomonas mobilis ZM4 gnlΔ, were investigated via growth inhibitory assay and biotransformation of glucose and fructose into gluconolactone and sorbitol, respectively. The results of ethanol fermentation studies performed in the presence of high concentration of glucose (>200 g l-1) under fermentative or aerobic conditions indicated that a significant reduction of volumetric ethanol productivity from the strain of ZM4 gnlΔ was noticeable due to the reduced rates of specific growth, sugar uptake, and biomass yield as compared with those of the parental strain ZM4. The biotransformation prepared at pH 6.0 using the permeabilized cell indicated that gluconic acid from ZM4 gnlΔ was still produced as a major product (67 g l-1) together with sorbitol (65 g l-1) rather than gluconolactone after 24 h. Only small amount of gluconolactone was transiently overproduced up to 9 g l-1, but at the end of biotransformation, all gluconolactone were oxidized into gluconic acid. This indicated that autolysis of gluconolactone at the pH led to such results despite under gluconolactonase inactivation conditions. The physiological characteristics of ZM4 gnlΔ was further investigated under various stress conditions, including suboptimal pH (3.5~6.0), temperature (25~40 °C), and presence of growth inhibitory molecules including hydrogen peroxide, ethanol, acetic acid, furfural, and so forth. The results indicated that ZM4 gnlΔ was more susceptible at high glucose concentration, low pH of 3.5, and high temperature of 40 °C and in the presence of 4 mM H2O2 comparing with ZM4. Therefore, the results were evident that gluconolactonase in Z. mobilis contributed to industrial robustness and anti-stress regulation.

The effect of humidified heated breathing circuit on core body temperature in perioperative hypothermia during thyroid surgery.

Purpose: During general anesthesia, human body easily reaches a hypothermic state, which is mainly caused by heat redistribution. Most studies suggested that humidified heated breathing circuits (HHBC) have little influence on maintenance of the core temperature during early phase of anesthesia. This study was aimed at examining heat preservation effect with HHBC in case of undergoing surgery with less exposure of surgical fields and short surgical duration. Methods: Patients aged 19 to 70 yr - old, ASA-PS I or II who were scheduled for elective thyroidectomy were assigned and divided to the group using HHBC (G1) and the group using conventional circuit (G2) by random allocation. During operation, core, skin, and room temperatures were measured every 5minutes by specific thermometer. Results: G1 was decreased by a lesser extent than G2 in core temperature, apparently higher at 30 and 60 minutes after induction. Skin and room temperatures showed no differences between the two groups (p>0.05). Consequently, we confirmed HHBC efficiently prevented a decrease in core temperature during early period in small operation which has difficulty in preparing warming devices or environments were not usually considered. Conclusions: This study showed that HHBC influences heat redistribution in early period of operation and can lessen the magnitude of the decrease in core body temperature. Therefore, it can be applied efficiently for other active warming devices in mild hypothermia.

The Halophilic Bacterium Paracoccus haeundaensis for the Production of Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) from Single Carbon Sources.

The study objective was to evaluate the potential production of polyhydroxyalkanoates (PHAs), a biodegradable plastic material, by Paracoccus haeundaensis for which PHA production has never been reported. To identify the most effective nitrogen-limited culture conditions for PHAs production from this bacterium, batch fermentation using glucose concentrations ranging from 4 g l-1 to 20 g l-1 with a fixed ammonium concentration of 0.5 g l-1 was carried out at 30°C and pH 8.0. A glucose supplement of 12 g l-1 produced the highest PHA concentration (1.6 g l-1) and PHA content (0.63 g g-1) thereby identifying the optimal condition for PHA production from this bacterium. Gas chromatography-mass spectrometry analysis suggests that P. haeundaensis mostly produced copolymer types of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] from glucose concentrations at 12 g l-1 or higher under the nitrogen-limited conditions. When several other single carbon sources were evaluated for the most efficient PHA production, fructose provided the highest biomass (2.8 g l-1), and PHAs (1.29 g l-1) concentrations. Results indicated that this bacterium mostly produced the copolymers P(3HB-co-3HV) from single carbon sources composing a range of 93-98% of 3-hydroxybutyrate and 2-7% of 3-hydroxyvalerate, whereas mannose-supplemented conditions produced the only homopolymer type of P(3HB). However, when propionic acid as a secondary carbon source were supplemented into the media, P. haeundaensis produced the copolymer P(3HB-co-3HV), composed of a 50% maximum monomeric unit of 3-hydroxyvaleric acid (3HV). However, as the concentration of propionic acid increased, cell biomass and PHAs concentrations substantially decreased due to cell toxicity.

Analgesic Effect of SKI306X on Chronic Postischemic Pain and Spinal Nerve Ligation-Induced Neuropathic Pain in Mice.

Neuropathic pain (NP) results from lesions or diseases affecting the peripheral or central somatosensory system. However, there are currently no drugs that are particularly effective in treating this condition. SKI306X is a blend of purified extracts of three oriental herbs (Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris) commonly used to treat osteoarthritis for their chondroprotective effects. Chronic postischemic pain (CPIP) and spinal nerve ligation (SNL) models were created by binding the upper left ankle of mice with an O-ring for 3 h and ligating the L5 spinal nerve, respectively. Mice with allodynia were injected intraperitoneally with 0.9% normal saline (NS group) or different doses (25, 50, or 100 mg/kg) of SKI306X (SKI groups). We assessed allodynia using von Frey filaments before injection and 30, 60, 90, 120, 180, and 240 min and 24 h after injection to confirm the antiallodynic effect of SKI306X. We also measured glial fibrillary acidic protein (GFAP) levels in the spinal cord and dorsal root ganglia to confirm the change of SKI306X administration. Both models exhibited significant mechanical allodynia. The intraperitoneal injection of SKI306X significantly increased the paw withdrawal threshold in a dose-dependent manner, as the paw withdrawal threshold was significantly increased after SKI306X administration compared with at baseline or after NS administration. GFAP levels in the SKI group decreased significantly (p < 0.05). Intraperitoneal administration of SKI306X dose-dependently attenuated mechanical allodynia and decreased GFAP levels, suggesting that GFAP is involved in the antiallodynic effect of SKI306X in mice with CPIP and SNL-induced NP.

Synergetic sustainability enhancement via current biofuel infrastructure: waste-to-energy concept for biodiesel production.

The concept of waste-to-energy (WtE) with regards to the utilization of byproducts from the bioethanol industry (e.g., distiller's dried grain with solubles: DDGS) was employed to enhance the renewability of biodiesel, which would be an initiative stage of a biorefinery due to the conjunction between bioethanol and biodiesel. For example, DDGS is a strong candidate for use as a biodiesel feedstock due to the tremendous amount that is regularly generated. On the basis of an estimation of possible lipid recovery from DDGS, ∼30% of the biodiesel feedstock demand in 2010 could be supported by the total DDGS generation in the same year. Considering the future expansion of the bioethanol industry up to 2020, the possible lipid recovery from DDGS would provide more than 6 times the biodiesel feedstock demand in 2010. In order to enhance the renewability of biodiesel, the transformation of lipid extracted from DDGS into fatty acid ethyl ester (FAEE) via a noncatalytic transesterification reaction under ambient pressure was investigated in this work. The newly introduced method reported here enables the combination of the esterification of free fatty acids (FFAs) and the transesterification of triglycerides into a single step. This was achieved in the presence of a porous material (i.e., charcoal), and the optimal conditions for transformation into biodiesel via this noncatalytic method were assessed at the fundamental level.

Effect of carbon dioxide on the thermal degradation of lignocellulosic biomass.

Using biomass as a renewable energy source via currently available thermochemical processes (i.e., pyrolysis and gasification) is environmentally advantageous owing to its intrinsic carbon neutrality. Developing methodologies to enhance the thermal efficiency of these proven technologies is therefore imperative. This study aimed to investigate the use of CO2 as a reaction medium to increase not only thermal efficiency but also environmental benefit. The influence of CO2 on thermochemical processes at a fundamental level was experimentally validated with the main constituents of biomass (i.e., cellulose and xylan) to avoid complexities arising from the heterogeneous matrix of biomass. For instance, gaseous products including H2, CH4, and CO were substantially enhanced in the presence of CO2 because CO2 expedited thermal cracking behavior (i.e., 200-1000%). This behavior was then universally observed in our case study with real biomass (i.e., corn stover) during pyrolysis and steam gasification. However, further study is urgently needed to optimize these experimental findings.

Biodiesel production from sewage sludge: new paradigm for mining energy from municipal hazardous material.

This work demonstrates that the production of biodiesel using the lipids extracted from sewage sludge (SS) could be economically feasible because of the remarkably high yield of oil and low cost of this feedstock, as compared to conventional biodiesel feedstocks. The yield of oil from SS, 980,000 L ha(-1) year(-1), is superior to those from microalgal and soybean oils, 446 and 2200 L ha(-1) year(-1), respectively. According to the case study of South Korea, the price of the lipids extracted from SS was approximately $0.03 L(-1) (USD), which is lower than those of all current biodiesel feedstocks. This work also highlights the insight of a novel methodology for transforming lipids containing high amounts of free fatty acids (FFAs) to biodiesel using a thermochemical process under ambient pressure in a continuous flow system. This allowed the combination of esterification of FFAs and transesterification of triglycerides into a single noncatalytic process, which led to a 98.5% ± 0.5% conversion efficiency to FAME (fatty acid methyl ester) within 1 min in a temperature range of 350-500 °C. The new process for converting the lipids extracted from SS shows high potential to achieve a major breakthrough in minimizing the cost of biodiesel production owing to its simplicity and technical advantages, as well as environmental benefits.

Genome-wide transcriptomic analysis of a flocculent strain of Zymomonas mobilis.

ZM401, a flocculent mutant strain of Zymomonas mobilis ZM4 was studied using genome-wide transcriptomic analysis for evidence related to phenotypic changes associated with its cell-cell attachment behaviour. Batch fermentation studies with ZM401 and its parent strain ZM4 demonstrated that similar ethanol yields and productivities could be achieved with both strains indicating the potential of the flocculent strains for cost-effective cell biomass recycling with resultant high ethanol volumetric productivities. The results showed that twofold or greater differential expression occurred for 26 genes of ZM401 when compared to those of ZM4. Among these, significant over-expression was evident for the genes ZMO1083 and ZMO1084 which are associated with bacterial cellulose synthesis, while reduced expression was found for ZMO0614, ZMO0613, and ZMO0635 which are all associated with synthesis of flagella-related proteins. Both enhanced cellulose production and reduced flagella activity are likely to facilitate more stable flocculent behaviour in ZM401. From comparative DNA sequence analysis of these 26 genes, only one single point mutation was identified. This occurred at the amino acid position A525V of ZMO1055 which encodes for diguanyl cyclase/phosphoesterase which may be related to cell motility and cellulose synthesis in Z. mobilis.

Functional use of CO2 to mitigate the formation of bisphenol A in catalytic pyrolysis of polycarbonate.

The growing consumption of plastic materials has increased hazardous threats to all environmental media, since current plastic waste management methods release microplastics and toxic chemicals. As such, massive generation of plastic derived pollutants leads to significant public health and environmental problems. In this work, an environmentally sound method for valorization of plastic waste is suggested. In detail, pyrolysis of polycarbonate-containing plastic waste such as automotive headlight housing (AHH) was carried out using CO2 as a co-reactant. AHH was chosen as it discharges bisphenol A (BPA) and aromatic compounds. Under CO2 condition, emissions of BPA and its derivatives were suppressed by 14.5% due to gas phase reactions (GPRs) with CO2. Nevertheless, reaction kinetics for GPRs was not significant. To impart the GPRs, catalytic pyrolysis was done using Ni and Co-based catalysts. During catalytic pyrolysis, syngas production was more than tenfold up comparing to pyrolysis without catalyst. The expedited GPRs over catalysts resulted in the enhanced syngas formation. Total concentration of the toxic chemicals from CO2-assisted catalytic pyrolysis of AHH decreased by 86.1% and 66.7% over Ni and Co catalysts, comparing to those from N2 environment.

Vitamin B12 biosynthesis gene diversity in the Ross Sea: the identification of a new group of putative polar B12 biosynthesizers.

Vitamin B12, a cobalt-containing micronutrient, has been shown to limit phytoplankton growth in the Ross Sea of the Southern Ocean. However, B12 biosynthesis potential in this environment remains uncharacterized. Select bacteria and archaea synthesize B12 while many phytoplankton require it for growth. Low ratios of bacterial biomass production to primary productivity and high concentrations of labile cobalt in Antarctic surface water suggest that factors controlling bacterial growth rather than cobalt availability may determine vitamin production rates here. In order to assess B12 biosynthesis potential, degenerate polymerase chain reaction primers were designed to target the genetic locus cbiA/cobB, encoding cobyrinic acid a,c-diamide synthase, a B12 biosynthesis protein. Sequencing the DNA compliment of Ross Sea 16S rRNA (see Supporting information) allowed targeting of cbiA/cobB probes to dominant bacterial groups. CbiA/cobB DNA sequences were successfully identified in clone libraries from the Ross Sea. To our knowledge, this study represents the first targeted molecular characterization of environmental B12 biosynthesis potential. A newly identified group of cbiA/cobB sequences dominated the diversity of the sequences retrieved; their expression was confirmed via mass spectrometry-based peptide detection. These sequences seem to have originated from a previously undescribed group of bacteria that could dominate the B12 biosynthesizing community in polar systems.

Assessment of salinity-induced photorespiratory glycolate metabolism in Anabaena sp. PCC 7120.

This paper reports an investigation of salinity-induced glycolate metabolism in the cyanobacterium Anabaena sp. PCC 7120 (hereafter Anabaena PCC 7120). Quantitative analysis of transcripts for the photosynthesis-associated genes encoding ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), phosphoribulokinase and transketolase, as well as those involved in glycolate metabolism (phosphoglycolate phosphatase, glycolate oxidase, alanine-glyoxylate aminotransferase and serine hydroxymethyltransferase) was performed. The expression of all investigated photosynthesis-associated genes except Rubisco was downregulated after 24 h NaCl treatment. However, under the same conditions, the transcripts encoding enzymes involved in glycolate metabolism were overexpressed. This was further confirmed by the quantitative analysis of the intermediates involved in glycolate metabolism. The intracellular levels of organic acids (glyceric, glycolic and glyoxylic acids) and amino acids (glycine and serine) were elevated in salt-treated cells as compared to those in the control cells. Transcriptional inhibition of photosynthesis-associated genes, and upregulation of genes and enhanced synthesis of intermediates associated with glycolate metabolism, indicate the occurrence of this photorespiratory metabolic pathway metabolism in Anabaena PCC 7120 under salt stress.

DNA restriction-modification systems in the ethanologen, Zymomonas mobilis ZM4.

To better understand the DNA restriction-modification (R-M) systems for more amenable strain development of the alternative industrial ethanologen, Zymomonas mobilis, three gene knockout mutants were constructed. The gene knockout mutants were tested for their DNA restriction activities by the determination of transformation efficiency using methylated and unmethylated foreign plasmid DNAs. Inactivation of a putative mrr gene encoded by ZMO0028 (zmrr) resulted in a 60-fold increase in the transformation efficiency when unmethylated plasmid DNA was used. This indicated that the putative mrr gene may serve as a type IV restriction-modification system in Z. mobilis ZM4. To assign the function of a putative type I DNA methyltransferase encoded by ZMO1933 (putative S subunit) and ZMO1934 (putative M subunit), the putative S subunit was inactivated. The gene inactivation of ZMO1933 resulted in a 30-fold increase in the transformation efficiency when methylated plasmid DNA was introduced, indicating that the putative S subunit possibly serves as a part of functional type I R-M system(s). Growth studies performed on the mutant strains indicate inactivation of the type I S subunit resulted in a lower maximum specific glucose consumption rate and biomass yield, while inactivation of the type IV Zmrr had the opposite effect, with an increase in the maximum specific growth rate and biomass yield.

Valorization of aflatoxin contaminated peanut into biodiesel through non-catalytic transesterification.

Aflatoxins (AFs) are the extremely hazardous metabolites (carcinogens) that are sporadically observed in crops, and these toxic chemicals are indeed lethal to the health of living organisms including human beings. Thus, AF contaminated food waste needs to be disposed as an environmentally benign way, not releasing it into the environment. This study offered a sustainable disposal and valorization platform for AF contaminated food. Peanut was used as a model food waste, because AF is readily appeared in the peanut during its harvesting, cultivation, storage, transportation process. As the valorization platform, non-catalytic transesterification of AF contaminated peanut was employed to convert it to biodiesel (BD). From the process, lipid in AF contaminated peanut is converted into BD (95.2 wt% yield) at 365°C for 1 min. Since the boiling points of BD and AF are significantly different, this process could also resolve the separation problem of AF (180 °C) from BD (≥ 330 °C) during the transesterification reaction. As a comparison study, alkali-catalyzed reaction was done. The alkali-catalyzed one required a pretreatment process to extract peanut oil for transesterification. The highest yield was 67.8 wt% yield after 6 h of reaction at 65 °C.

Valorization of carbon dioxide and waste (Derived from the site of Eutrophication) into syngas using a catalytic thermo-chemical platform.

Global warming increases a chance of eutrophication, and such fact offers that unhygienic organic waste materials (OWMs) in water must be treated. Hence, this study laid emphasis on the thermal-chemical (pyrolysis) process to establish a rapid valorization platform for OWMs. Indeed, OWMs were collected from the eutrophication site, and OWMs were mainly comprised of lignocellulosic biomass, microalgae (cyanobacteria) and the diverse types of bacteria (commonly observed from livestock waste). In an attempt to offer more sustainable valorization route for OWMs, CO2 was used as a raw material in pyrolysis process. From the CO2-assisted pyrolysis, the conversion of CO2 and OWMs into gaseous fuel (CO) was observed. A cheap Ni-based catalyst was used in pyrolysis of OWMs as a strategic practice to promote conversion of CO2 into CO. Indeed, syngas production (38 %) was enhanced from catalytic pyrolysis over Ni/SiO2 under CO2 condition as compared to inert condition (N2).

Pikeienuella piscinae gen. nov., sp. nov., a novel genus in the family Rhodobacteraceae.

A novel bacterium, designated strain RR4-56T, was isolated from a biofilter of a seawater recirculating aquaculture system. The 16S rRNA gene sequence analysis showed that the isolate was closely related to Halovulum dunhuangense YYQ-30T (92.6%), Albimonas donghaensis DS2T (91.3%), Pontivivens insulae GYSW-23T (91.3%), and Monaibacterium marinum C7T (90.9%), belonging to the family Rhodobacteraceae. The strain was aerobic, Gram-negative, rod-shaped, oxidase-positive, and catalase-negative. Its optimum temperature, pH, and salinity for growth were 25-30°C, pH 8.5, and 2-3% NaCl (w/v), respectively. Its growth occurred at 15-35°C, pH 5.0-9.5, and 0-7% NaCl (w/v). It contained ubiquinone-10 (Q-10), a respiratory quinone, and the major cellular fatty acids were 11-methyl C18:1 ω7c (31.9%), C18:1 ω6c (30.4%), and C19:0 cyclo ω8c (16.1%). The polar lipids present in the strain were phosphatidylglycerol, an unidentified phospholipid, and an unidentified aminolipid. The strain had one 4,373,045 bp circular chromosome with G + C contents of 65.9 mol% including 4,169 genes, 4,118 coding sequences (CDSs), 3 rRNAs, and 45 tRNAs. Genome annotation predicted some gene clusters related to the degradation of several types of organic matter such as protocatechuate, catechol, and phthalate. Based on the polyphasic characteristics, RR4-56T represents a novel genus and species in the family Rhodobacteraceae, for which the name Pikeienuella piscinae gen. nov., sp. nov. was proposed. The type strain is RR4-56T (= KCTC 52648T = DSM 107918T).

Biodiesel synthesis from swine manure.

This study demonstrates that the biodiesel (BD) from swine manure (SM) could be a promising way for large scale generation of biofuel. Also, the economic and environmental benefits of SM derived BD were evaluated. Transesterification of lipid contents extracted from the collected SM had low BD yield (14.2 wt%) using H2SO4 catalyst due to high acid value and impurities. However, thermo-chemical non-catalytic transesterification with a porous material showed 94.7 wt% yield of BD from the lipid in SM. Considering the current population of swine, the annual production of BD from SM was estimated. The SM derived BD could cover 19.7 and 46.8 wt% of BD currently produced in both Korea and the USA with the economic benefits of up to $96 million and $2.1 billion, respectively. The proposed approach also can save vast arable lands needed to cultivate oil-bearing feedstocks for BD production.

Effect of biochars pyrolyzed in N2 and CO2, and feedstock on microbial community in metal(loid)s contaminated soils.

Little is known about the effects of applying amendments on soil for immobilizing metal(loid)s on the soil microbial community. Alterations in the microbial community were examined after incubation of treated contaminated soils. One soil was contaminated with Pb and As, a second soil with Cd and Zn. Red pepper stalk (RPS) and biochars produced from RPS in either N2 atmosphere (RPSN) or CO2 atmosphere (RPSC) were applied at a rate of 2.5% to the two soils and incubated for 30 days. Bacterial communities of control and treated soils were characterized by sequencing 16S rRNA genes using the Illumina MiSeq sequencing. In both soils, bacterial richness increased in the amended soils, though somewhat differently between the treatments. Evenness values decreased significantly, and the final overall diversities were reduced. The neutralization of pH, reduced available concentrations of Pb or Cd, and supplementation of available carbon and surface area could be possible factors affecting the community changes. Biochar amendments caused the soil bacterial communities to become more similar than those in the not amended soils. The bacterial community structures at the phylum and genus levels showed that amendment addition might restore the normal bacterial community of soils, and cause soil bacterial communities in contaminated soils to normalize and stabilize.

Production of bioplastic through food waste valorization.

The tremendous amount of food waste from diverse sources is an environmental burden if disposed of inappropriately. Thus, implementation of a biorefinery platform for food waste is an ideal option to pursue (e.g., production of value-added products while reducing the volume of waste). The adoption of such a process is expected to reduce the production cost of biodegradable plastics (e.g., compared to conventional routes of production using overpriced pure substrates (e.g., glucose)). This review focuses on current technologies for the production of polyhydroxyalkanoates (PHA) from food waste. Technical details were also described to offer clear insights into diverse pretreatments for preparation of raw materials for the actual production of bioplastic (from food wastes). In this respect, particular attention was paid to fermentation technologies based on pure and mixed cultures. A clear description on the chemical modification of starch, cellulose, chitin, and caprolactone is also provided with a number of case studies (covering PHA-based products) along with a discussion on the prospects of food waste valorization approaches and their economic/technical viability.

Biosynthetic intermediate analysis and functional homology reveal a saxitoxin gene cluster in cyanobacteria.

Saxitoxin (STX) and its analogues cause the paralytic shellfish poisoning (PSP) syndrome, which afflicts human health and impacts coastal shellfish economies worldwide. PSP toxins are unique alkaloids, being produced by both prokaryotes and eukaryotes. Here we describe a candidate PSP toxin biosynthesis gene cluster (sxt) from Cylindrospermopsis raciborskii T3. The saxitoxin biosynthetic pathway is encoded by more than 35 kb, and comparative sequence analysis assigns 30 catalytic functions to 26 proteins. STX biosynthesis is initiated with arginine, S-adenosylmethionine, and acetate by a new type of polyketide synthase, which can putatively perform a methylation of acetate, and a Claisen condensation reaction between propionate and arginine. Further steps involve enzymes catalyzing three heterocyclizations and various tailoring reactions that result in the numerous isoforms of saxitoxin. In the absence of a gene transfer system in these microorganisms, we have revised the description of the known STX biosynthetic pathway, with in silico functional inferences based on sxt open reading frames combined with liquid chromatography-tandem mass spectrometry analysis of the biosynthetic intermediates. Our results indicate the evolutionary origin for the production of PSP toxins in an ancestral cyanobacterium with genetic contributions from diverse phylogenetic lineages of bacteria and provide a quantum addition to the catalytic collective available for future combinatorial biosyntheses. The distribution of these genes also supports the idea of the involvement of this gene cluster in STX production in various cyanobacteria.