Orientalis Strains Research Articles

A transcriptomic atlas of acute stress response to low pH in multiple Issatchenkia orientalis strains.

Issatchenkia orientalis is a promising industrial chassis to produce biofuels and bioproducts due to its high tolerance to multiple environmental stresses such as low pH, heat, and other chemicals otherwise toxic for the most widely used microbes. Yet, little is known about specific mechanisms of such tolerance in this organism, hindering our ability to engineer this species to produce valuable biochemicals. Here, we report a comprehensive study of the mechanisms of acidic tolerance in this species via transcriptome profiling across variable pH for 12 different strains with different phenotypes. We found multiple regulatory mechanisms involved in tolerance to low pH in different strains of I. orientalis, marking potential targets for future gene editing and perturbation experiments.

Identification of a unique conserved region from a kinetoplastid genome of Leishmania orientalis (formerly named Leishmania siamensis) strain PCM2 in Thailand

Mitochondrial DNAs (mtDNAs) appear in almost all eukaryotic species and are useful molecular markers for phylogenetic studies and species identification. Kinetoplast DNAs (kDNAs) are structurally complex circular mtDNA networks in kinetoplastids, divided into maxicircles and minicircles. Despite several kDNAs of many Leishmania species being examined, the kDNAs of the new species, Leishmania orientalis (formerly named Leishmania siamensis) strain PCM2, have not been explored. This study aimed to investigate the maxicircle and minicircle DNAs of L. orientalis strain PCM2 using hybrid genome sequencing technologies and bioinformatic analyses. The kDNA sequences were isolated and assembled using the SPAdes hybrid assembler from the Illumina short-read and PacBio long-read data. Circular contigs of the maxicircle and minicircle DNAs were reconstructed and confirmed by BLASTn and rKOMICs programs. The kDNA genome was annotated by BLASTn before the genome comparison and phylogenetic analysis by progressiveMauve, MAFFT, and MEGA programs. The maxicircle of L. orientalis strain PCM2 (18,215 bp) showed 99.92% similarity and gene arrangement to Leishmania enriettii strain LEM3045 maxicircle with variation in the 12s rRNA gene and divergent region. Phylogenetics of the whole sequence, coding regions, divergent regions, and 12s rRNA gene also confirmed this relationship and subgenera separation. The identified 105 classes of minicircles (402–1177 bp) were clustered monophyletically and related to the Leishmania donovani minicircles. The kinetoplast maxicircle and minicircle DNAs of L. orientalis strain PCM2 contained a unique conserved region potentially useful for specific diagnosis of L. orientalis and further exploration of this parasite population genetics in Thailand and related regions.

Application of Clostridium butyricum, Rummeliibacillus suwonensis, and Issatchenkia orientalis for Nongxiangxing baijiu fermentation: Improves the microbial communities and flavor of upper fermented grain

Ethyl hexanoate and ethyl butyrate are essential to the flavor compounds in Nongxiangxing baijiu, but low levels of these two esters in upper fermented grains (FG) decreases the quality of upper distilled baijiu, representing the main challenge in Nongxiangxing baijiu production. This paper enhanced fermentation by inoculating functional Clostridium butyricum, Rummeliibacillus suwonensis, and Issatchenkia orientalis strains into upper FG. The results showed that the ethyl butyrate content in the upper FG increased significantly and the content of ethyl hexanoate did improve from the results of many determinations. High-throughput sequencing indicated that the dominant phyla in the FG were Firmicutes, Actinobacteriota, Proteobacteria, Ascomycota, and Basidiomycota. The canonical correspondence analysis (CCA) and person correlation network revealed the relationship between the microbial community, physicochemical environment, and flavor compounds. The temperature, oxygen, and acidity were closely related to the microbial community, while most flavor compounds were positively correlated with Caldicoprobacter, Caproiciproducens, Delftia, Hydrogenispora, Thermoactinomyces, Issatchenkia Bacillus, and Aspergillus. These results helped improve the quality of Nongxiangxing baijiu.

Cas9-Based Metabolic Engineering of Issatchenkia orientalis for Enhanced Utilization of Cellulosic Hydrolysates.

Issatchenkia orientalis, exhibiting high tolerance against harsh environmental conditions, is a promising metabolic engineering host for producing fuels and chemicals from cellulosic hydrolysates containing fermentation inhibitors under acidic conditions. Although genetic tools for I. orientalis exist, they require auxotrophic mutants so that the selection of a host strain is limited. We developed a drug resistance gene (cloNAT)-based genome-editing method for engineering any I. orientalis strains and engineered I. orientalis strains isolated from various sources for xylose fermentation. Specifically, xylose reductase, xylitol dehydrogenase, and xylulokinase from Scheffersomyces stipitis were integrated into an intended chromosomal locus in four I. orientalis strains (SD108, IO21, IO45, and IO46) through Cas9-based genome editing. The resulting strains (SD108X, IO21X, IO45X, and IO46X) efficiently produced ethanol from cellulosic and hemicellulosic hydrolysates even though the pH adjustment and nitrogen source were not provided. As they presented different fermenting capacities, selection of a host I. orientalis strain was crucial for producing fuels and chemicals using cellulosic hydrolysates.

Making the Most of Its Short Reads: A Bioinformatics Workflow for Analysing the Short-Read-Only Data of Leishmania orientalis (Formerly Named Leishmania siamensis) Isolate PCM2 in Thailand.

Simple SummaryLeishmaniasis is a parasitic disease caused by flagellated protozoa of the genus Leishmania. Multiple genome sequencing platforms have been employed to complete Leishmania genomes at the expense of high cost. This study proposes an integrative bioinformatic workflow for assembling only the short-read data of Leishmania orientalis isolate PCM2 from Thailand and produce an acceptable-quality genome for further genomic analysis. This workflow gives extensive information required for identifying strain-specific markers and virulence-associated genes useful for drug and vaccine development before a more exhaustive and expensive investigation.Background: Leishmania orientalis (formerly named Leishmania siamensis) has been neglected for years in Thailand. The genomic study of L. orientalis has gained much attention recently after the release of the first high-quality reference genome of the isolate LSCM4. The integrative approach of multiple sequencing platforms for whole-genome sequencing has proven effective at the expense of considerably expensive costs. This study presents a preliminary bioinformatic workflow including the use of multi-step de novo assembly coupled with the reference-based assembly method to produce high-quality genomic drafts from the short-read Illumina sequence data of L. orientalis isolate PCM2. Results: The integrating multi-step de novo assembly by MEGAHIT and SPAdes with the reference-based method using the L. enriettii genome and salvaging the unmapped reads resulted in the 30.27 Mb genomic draft of L. orientalis isolate PCM2 with 3367 contigs and 8887 predicted genes. The results from the integrated approach showed the best integrity, coverage, and contig alignment when compared to the genome of L. orientalis isolate LSCM4 collected from the northern province of Thailand. Similar patterns of gene ratios and frequency were observed from the GO biological process annotation. Fifty GO terms were assigned to the assembled genomes, and 23 of these (accounting for 61.6% of the annotated genes) showed higher gene counts and ratios when results from our workflow were compared to those of the LSCM4 isolate. Conclusions: These results indicated that our proposed bioinformatic workflow produced an acceptable-quality genome of L. orientalis strain PCM2 for functional genomic analysis, maximising the usage of the short-read data. This workflow would give extensive information required for identifying strain-specific markers and virulence-associated genes useful for drug and vaccine development before a more exhaustive and expensive investigation.

Theileria orientalis outbreak in an organized cattle breeding farm.

Theileriosis is an important tick borne disease of cattle caused by a haemoprotozoan of genus Theileria. Clinical bovine theileriosis is mainly caused by T. annulata or T. parva but the clinical disease due to T. orientalis is rare. T. orientalis mainly infect RBCs and causes "Oriental theileriosis" or Theileria-associated bovine anaemia in cattle and other livestock species. Two genotypes of T. orientalis (Chitose and Ikeda) are reported to cause severe disease in some countries. In this report, a spontaneous outbreak of Oriental theileriosis was studied in an organized Holstein-Friesian cattle breeding farm situated in the south-eastern Himalayan ranges of Himachal Pradesh State of India. Animal blood and tick samples were tested using cytological and PCR techniques. The disease episode occurred in a protracted manner spanning over 10 to 12 months and association of T. orientalis was confirmed in 93.3% of the blood and 21.7% of Rhipicephalus microplus (tick) samples. No other tick borne pathogen was detected except Anaplasma marginale in two blood samples. Haematological profiling of infected cattle showed characteristic indicators of anaemia like haemoblobin, RBC count, haematocrit value and mean corpuscular volume at either lower than normal or near the lower normal range. The prevailing persistent anaemic changes led to more severe clinical manifestations like abortion and joint inflammation. The detected T. orientalis strains and ticks species were further confirmed by nucleotide sequence analysis of 18S rRNA and 16S rRNA genes. Phylogenetically, T. orientalis strains showed clustering with other reported strains of T. orientalis from the surrounding regions. This first report of clinical Oriental theileriosis from India emphasises the importance of T. orientalis as an emerging tick borne pathogen and role of widely prevalent ticks species in disease transmission and their impact on livestock production.

Ethanol Production from Wheat Straw Hydrolysate by Issatchenkia Orientalis Isolated from Waste Cooking Oil

The interest in using non-conventional yeasts to produce value-added compounds from low cost substrates, such as lignocellulosic materials, has increased in recent years. Setting out to discover novel microbial strains that can be used in biorefineries, an Issatchenkia orientalis strain was isolated from waste cooking oil (WCO) and its capability to produce ethanol from wheat straw hydrolysate (WSHL) was analyzed. As with previously isolated I. orientalis strains, WCO-isolated I. orientalis KJ27-7 is thermotolerant. It grows well at elevated temperatures up to 42 °C. Furthermore, spot drop tests showed that it is tolerant to various chemical fermentation inhibitors that are derived from the pre-treatment of lignocellulosic materials. I. orientalis KJ27-7 is particularly tolerant to acetic acid (up to 75 mM) and tolerates 10 mM formic acid, 5 mM furfural and 10 mM hydroxymethylfurfural. Important for biotechnological cellulosic ethanol production, I. orientalis KJ27-7 grows well on plates containing up to 10% ethanol and media containing up to 90% WSHL. As observed in shake flask fermentations, the specific ethanol productivity correlates with WSHL concentrations. In 90% WSHL media, I. orientalis KJ27-7 produced 10.3 g L−1 ethanol within 24 h. This corresponds to a product yield of 0.50 g g−1 glucose (97% of the theoretical maximum) and a volumetric productivity of 0.43 g L−1 h−1. Therefore, I. orientalis KJ27-7 is an efficient producer of lignocellulosic ethanol from WSHL.

Metschnikowia miensis f.a., sp. nov., isolated from flowers in Mie prefecture, Japan.

Four yeast strains (RIFY 10001T, RIFY 10002, RIFY 10003, and RIFY 10004) were isolated from flowers growing in fields of mustard and broad beans in Japan. Ascospore formation was not observed. Sequence analysis of the D1/D2 domain of the large subunit ribosomal RNA (LSU rRNA) gene of the four strains indicated that they belong to the genus Metschnikowia and are closely related to Metschnikowia hawaiiana strain CBS 9146T and Metschnikowia orientalis strain CBS 10331T. The D1/D2 domain of the LSU rRNA gene and internal transcribed spacer regions of strain RIFY 10001T were 85.7% identical to those of M. hawaiiana strain CBS 9146T. All four strains were distinguished from the M. hawaiiana strain CBS 9146T by their inability to ferment glucose. Hence, these four strains are novel species and were named as Metschnikowia miensis (holotype: RIFY 10001T; isotypes: NBRC 112445T = CBS 14749T).

The Influence of Selected Plant Essential Oils on Morphological and Physiological Characteristics in Pseudomonas Orientalis.

The aim of this work was to estimate the antibacterial activity of selected essential oils on Pseudomonas orientalis strains isolated from foods. An attempt was also made to identify the mechanisms of the action of the plant oils. Classical methods of assessment of the effectiveness of antimicrobial activity of oils were linked with flow cytometry. It was observed that bergamot, lemongrass, bitter orange, juniper, and black pepper oils have bacteriostatic effect against P. orientalis P49. P. orientalis P110 is sensitive to lime, lemongrass, juniper, rosemary, and black pepper oils. Additionally, plant oils with biostatic effect on P. orientalis limited the intracellular metabolic activity of cells; this was closely linked with the ability of plant oils’ bioactive components to interact with bacteria cell membrane, causing the release of membrane proteins. As a result, the selective permeability of the cell membranes were damaged and the bacterial shape was transformed to coccoid in form.

Influence of sub-inhibitory concentration of selected plant essential oils on the physical and biochemical properties of Pseudomonas orientalis

AbstractPlant essential oils have bacteriostatic activity in very low concentrations, thus they can be used as natural preservative in food products. The aim of this work was to understand the inhibitory and non-bactericidal effects of essential oils in low concentrations. To meet this aim some physical and biochemical properties of two P. orientalis strains isolated from food probes were investigated under the influence of plant oils. Control probes - without oils - played an important role in our investigations because P. orientalis is not a well-known and described species. Thus, these results allow us to gain additional knowledge about bacteria from P. orientalis species. The obtained results are proof that under the influence of oils, bacteria lose their ability to move, change their morphology, and also reduce their metabolic activity. However, they do not die, and properties such as the ability to produce ammonia, the ability to production of indole from the amino acid tryptophan as well as the ability to assimilation of saccharides are maintained.

Alleviation of metal stress by Pseudomonas orientalis and Chaetomium cupreum strains and their effects on Eucalyptus globulus growth promotion

Pseudomonas orientalis and Chaetomium cupreum isolated from metal(oid)s-contaminated soil were characterized to evaluate their role in plant growth promotion and enhance the tolerance of Eucalyptus globulus to copper stress. We evaluated minimum inhibitory concentration and explored plant-promoting traits under copper stress. Moreover, changes in morphology and the accumulation of metals on the cell surface were determined by scanning electron microscopy images coupled with energy dispersive x-ray spectroscopy (SEM-EDX). The effect of inoculation with P. orientalis and C. cupreum on E. globulus plants established in soils contaminated with copper was determined through root and shoot biomass and antioxidant response. Both strains showed high tolerance to metal(oid)s, whereas phosphate solubilization was only detected in P. orientalis. Under copper stress, C. cupreum presented a higher production rate of siderophores and indole acetic acid (4.5 mm day−1, 15.042 μg mL−1). SEM images and EDX analysis reflected cellular changes in fungus and proved biosorption of copper. Inoculation of P. orientalis and C. cupreum on Eucalyptus globulus plants significantly increased plant growth and mitigated the toxic effects of copper. According to these results, both microorganisms contribute positively to stimulated plant growth and to decreasing stress caused by high concentrations of copper in soil, especially C. cupreum.

Endophytic Bacillus and Pseudomonas spp. Modulate Apple Shoot Growth, Cellular Redox Balance, and Protein Expression Under in Vitro Conditions.

Interactions between host plants and endophytic microorganisms play an important role in plant responses to pathogens and environmental stresses and have potential applications for plant stress management under in vitro conditions. We assessed the effect of endophytic bacteria on the growth and proliferation of domestic apple cv. Gala shoots in vitro. Further, a model apple cell suspension system was used to examine molecular events and protein expression patterns at an early stage of plant–endophyte interaction. Among the seven strains used in the study, Bacillus spp. strains Da_1, Da_4, and Da_5 and the Pseudomonas fluorescens strain Ga_1 promoted shoot growth and auxiliary shoot proliferation. In contrast, Bacillus sp. strain Oa_4, P. fluorescens strain Ga_3 and P. orientalis strain G_12 inhibited shoot development. In the cell suspension, the effects of the association between endophytic bacteria and plant cells were specific to each strain. Modulation of the cellular redox balance was monitored in the apple cells using a 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) probe, and strain-specific effects were observed that correlated with the in vitro shoot development results. Proteomic analysis revealed differences in protein expressions in apple cells co-cultivated with different Bacillus spp. strains that had contrasting effects on cellular redox balance and shoot development. The Bacillus sp. strain Da_4, which enhanced shoot development and oxidation of H2DCFDA, induced differential expression of proteins that are mainly involved in the defense response and regulation of oxidative stress. Meanwhile, treatment with Bacillus sp. strain Oa_4 led to strong upregulation of PLAT1, HSC70-1 and several other proteins involved in protein metabolism and cell development. Taken together, the results suggest that different cell signaling and response events at the early stage of the plant–endophyte interaction may be important for strain-dependent regulation of cellular redox balance and development of shoot phenotype.

Whole-Genome Sequence of Acetobacter orientalis Strain FAN1, Isolated from Caucasian Yogurt.

ABSTRACTIn traditional Caucasian yogurt, Acetobacter orientalis bacteria play important roles in the fermentation of milk in concert with Lactococcus bacteria. In this study, an A. orientalis strain, FAN1, was newly isolated from commercially available Caucasian yogurt, and its whole-genome sequence was determined, identifying two circular DNAs.

Selection and use of indigenous mixed starter cultures for mustard leaves fermentation and the improvement of cuocai characteristics

To improve the characteristics and safety of traditional cuocai, the selection and use of mixed starter cultures is important. This study aimed to isolate the main indigenous lactic acid bacteria (LABs) and yeast species from spontaneous fermented mustard leaves, and they were used as starter cultures to ferment mustard leaves. Five predominant LABs, namely Q-1 (Enterococcus faecalis), G-1 (Lactobacillus plantarum), G-3 (Lactobacillus pentosus), and G-2 and G-4 (Lactobacillus buchneri) strains, and the two yeast strains J-1 (Issatchenkia orientalis) and J-2 (Issatchenkia occidentalis) were isolated from cuocai. From the results, Lb. plantarum and I. orientalis strains exhibited strong growth ability, fast acidification, and potent low-pH tolerance. Therefore, they were chosen as the mixed starter cultures to ferment mustard leaves. During the process of culture-dependent fermented mustard leaves, the changes of acidity, total crude protein, and total sugar were similar to those of culture-independent fermented mustard leaves, while the nitrite content of culture-dependent fermented mustard leaves was significantly lower than that of culture-independent fermented cuocai. According to gas chromatography-mass spectrometry analysis, the main volatile flavor compounds of fermented mustard leaves were alcohols, aldehydes, esters, and sulfides. There was no significant difference of volatile flavor compounds between the processes of culture-dependent and culture-independent fermentations. Therefore, it was suggested that Lb. plantarum and I. orientalis strains could be used to ferment cuocai instead of the traditional ferment method to control the quality and shelf safety of cuocai. © 2017 Society of Chemical Industry.

A systematic study of the whole genome sequence of Amycolatopsis methanolica strain 239T provides an insight into its physiological and taxonomic properties which correlate with its position in the genus

The complete genome of methanol-utilizing Amycolatopsis methanolica strain 239T was generated, revealing a single 7,237,391 nucleotide circular chromosome with 7074 annotated protein-coding sequences (CDSs). Comparative analyses against the complete genome sequences of Amycolatopsis japonica strain MG417-CF17T, Amycolatopsis mediterranei strain U32 and Amycolatopsis orientalis strain HCCB10007 revealed a broad spectrum of genomic structures, including various genome sizes, core/quasi-core/non-core configurations and different kinds of episomes. Although polyketide synthase gene clusters were absent from the A. methanolica genome, 12 gene clusters related to the biosynthesis of other specialized (secondary) metabolites were identified. Complete pathways attributable to the facultative methylotrophic physiology of A. methanolica strain 239T, including both the mdo/mscR encoded methanol oxidation and the hps/hpi encoded formaldehyde assimilation via the ribulose monophosphate cycle, were identified together with evidence that the latter might be the result of horizontal gene transfer. Phylogenetic analyses based on 16S rDNA or orthologues of AMETH_3452, a novel actinobacterial class-specific conserved gene against 62 or 18 Amycolatopsis type strains, respectively, revealed three major phyletic lineages, namely the mesophilic or moderately thermophilic A. orientalis subclade (AOS), the mesophilic Amycolatopsis taiwanensis subclade (ATS) and the thermophilic A. methanolica subclade (AMS). The distinct growth temperatures of members of the subclades correlated with corresponding genetic variations in their encoded compatible solutes. This study shows the value of integrating conventional taxonomic with whole genome sequence data.

First identification of Francisella noatunensis subsp. orientalis causing mortality in Mexican tilapia Oreochromis spp.

Francisellosis, an emerging disease in tilapia Oreochromis spp., is caused by the facultative, intracellular bacterium Francisella noatunensis subsp. orientalis, which is present in various countries where tilapia farming is commercially important. We confirmed the presence of francisellosis in Mexican tilapia cultures in association with an outbreak during the second semester of 2012. Broodstock fish presented a mortality rate of approximately 40%, and disease was characterized by histologically classified granulomas, or whitish nodules, in different organs, mainly the spleen and kidney. Through DNA obtained from infected tissue and pure cultures in a cysteine heart medium supplemented with hemoglobin, F. noatunensis subsp. orientalis was initially confirmed through the amplification and analysis of the 16S rRNA gene and the internal transcribed spacer region. Phylogenetic analysis of these genes demonstrated close similarity with previously reported F. noatunensis subsp. orientalis sequences obtained from infected tilapia from various countries. The identification of this subspecies as the causative agent of the outbreak was confirmed using the iglC gene as a target sequence, which showed 99.5% identity to 2 F. noatunensis subsp. orientalis strains (Ethime-1 and Toba04). These findings represent the first documented occurrence of francisellosis in Mexican tilapia cultures, which highlights the importance of establishing preventative measures to minimize the spread of this disease within the Mexican aquaculture industry.

Whole-Genome Sequence of Francisella noatunensis subsp. orientalis Strain FNO01 Isolated from Diseased Nile Tilapia in Brazil.

This paper describes the complete genome sequence of Francisella noatunensis subsp. orientalis strain FNO01, which was isolated during the first outbreak of francisellosis in cultured Nile tilapia in Brazil. The genome is composed of a circular chromosome with 1,859,830 bp and a G+C content of ~32%.

Occurrence of Cellulose-Producing Gluconacetobacter spp. in Fruit Samples and Kombucha Tea, and Production of the Biopolymer.

Cellulose producing bacteria were isolated from fruit samples and kombucha tea (a fermented beverage) using CuSO4 solution in modified Watanabe and Yamanaka medium to inhibit yeasts and molds. Six bacterial strains showing cellulose production were isolated and identified by 16S rRNA gene sequencing as Gluconacetobacter xylinus strain DFBT, Ga. xylinus strain dfr-1, Gluconobacter oxydans strain dfr-2, G. oxydans strain dfr-3, Acetobacter orientalis strain dfr-4, and Gluconacetobacter intermedius strain dfr-5. All the cellulose-producing bacteria were checked for the cellulose yield. A potent cellulose-producing bacterium, i.e., Ga. xylinus strain DFBT based on yield (cellulose yield 5.6g/L) was selected for further studies. Cellulose was also produced in non- conventional media such as pineapple juice medium and hydrolysed corn starch medium. A very high yield of 9.1g/L cellulose was obtained in pineapple juice medium. Fourier transform infrared spectrometer (FT-IR) analysis of the bacterial cellulose showed the characteristic peaks. Soft cellulose with a very high water holding capacity was produced using limited aeration. Scanning electron microscopy (SEM) was used to analyze the surface characteristics of normal bacterial cellulose and soft cellulose. The structural analysis of the polymer was performed using (13)C solid-state nuclear magnetic resonance (NMR). More interfibrillar space was observed in the case of soft cellulose as compared to normal cellulose. This soft cellulose can find potential applications in the food industry as it can be swallowed easily without chewing.

Enzymatic saccharification of cassava residues and glucose inhibitory kinetics on β-glucosidase from Hypocrea orientalis.

Cassava residues are byproducts of the starch industry containing abundant cellulose for bioproduction of green fuel. To obtain maximum sugar yields from cassava residues, the optimal conditions for hydrolyzing the residues were determined using cellulase prepared from a novel Hypocrea orientalis strain. The optimal pH value and optimal temperature for the cellulase hydrolysis were 5.0 and 50 °C, respectively. The concentration of NaOH was determined to be 1% for pretreatment of cassava residues to gain enough soluble sugars suitably. The yield of released sugars was 10 mg/mL in the optimal conditions after 24 h of reaction, which was similar to that of bagasse and wheat grass. Inhibition kinetics of H. orientalis β-glucosidase (BG) by glucose was first studied using the progress-of-substrate-reaction method as described by Tsou (Tsou, C. L. Adv. Enzymol. Related Areas Mol. Biol. 1988, 61, 381-436), and the microscopic inhibition rate constants of glucose were determined. The results showed that glucose could inhibit BG reversibly and competitively. The rate constants of forward (k(+0)) and reverse (k(-0)) reaction were measured to be 4.88 × 10(-4) (mM·s)(-1) and 2.7 × 10(-4) s(-1), respectively. Meanwhile, the inhibition was more significant than that of L-glucose, D-mannose, D-galactose, D-aminoglucose, acetyl-D-glucose, and D-fructose. This work reveals how to increase sugar yields and reduce product inhibition during enzymatic saccharification of cellulose.

Functional characterization of Yersinia pestis aerobic glycerol metabolism

Yersinia pestis biovar Orientalis isolates have lost the capacity to ferment glycerol. Herein we provide experimental validation that a 93 bp in-frame deletion within the glpD gene encoding the glycerol-3-phosphate dehydrogenase present in all biovar Orientalis strains is sufficient to disrupt aerobic glycerol fermentation. Furthermore, the inability to ferment glycerol is often insured by a variety of additional mutations within the glpFKX operon which prevents glycerol internalization and conversion to glycerol-3-phosphate. The physiological impact of functional glpFKX in the presence of dysfunctional glpD was assessed. Results demonstrate no change in growth kinetics at 26 °C and 37 °C. Mutants deficient in glpD displayed decreased intracellular accumulation of glycerol-3-phosphate, a characterized inhibitor of cAMP receptor protein (CRP) activation. Since CRP is rigorously involved in global regulation Y. pestis virulence, we tested a possible influence of a single glpD mutation on virulence. Nonetheless, subcutaneous and intranasal murine challenge was not impacted by glycerol metabolism. As quantified by crystal violet assay, biofilm formation of the glpD-deficient KIM6+ mutant was mildly repressed; whereas, chromosomal restoration of glpD in CO92 resulted in a significant increase in biofilm formation.