Indole acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solanum lycopersicum

BackgroundEndophytic bacteria are ubiquitous in all plant species contributing in host plant's nutrient uptake and helping the host to improve its growth. Moringa peregrina which is a medicinal plant, growing in arid region of Arabia, was assessed for the presence of endophytic bacterial strains.ResultsPCR amplification and sequencing of 16S rRNA of bacterial endophytes revealed the 5 endophytic bacteria, in which 2 strains were from Sphingomonas sp.; 2 strains from Bacillus sp. and 1 from Methylobacterium genus. Among the endophytic bacterial strains, a strain of Bacillus subtilis LK14 has shown significant prospects in phosphate solubilization (clearing zone of 56.71 mm after 5 d), ACC deaminase (448.3 ± 2.91 nM α-ketobutyrate mg^- 1 h^- 1) and acid phosphatase activity (8.4 ± 1.2 nM mg^- 1 min^- 1). The endophytic bacteria were also assessed for their potential to produce indole-3-acetic acid (IAA). Among isolated strains, the initial spectrophotometry analysis showed significantly higher IAA production by Bacillus subtilis LK14. The diurnal production of IAA was quantified using multiple reactions monitoring method in UPLC/MS–MS. The analysis showed that LK14 produced the highest (8.7 μM) IAA on 14th d of growth. Looking at LK14 potentials, it was applied to Solanum lycopersicum, where it significantly increased the shoot and root biomass and chlorophyll (a and b) contents as compared to control plants.ConclusionThe study concludes that using endophytic bacterial strains can be bio-prospective for plant growth promotion, which might be an ideal strategy for improving growth of crops in marginal lands.     

Molecular cloning,expression, and immobilization of glutamate decarboxylase from Lactobacillus fermentum YS2

BackgroundGABA (γ-aminobutyric acid) is a four-carbon nonprotein amino acid that has hypotensive, diuretic, and tranquilizing properties. Glutamate decarboxylase (GAD) is the key enzyme to generate GABA. A simple and economical method of preparing and immobilizing GAD would be helpful for GABA production. In this study, the GAD from Lactobacillus fermentum YS2 was expressed under the control of a stress-inducible promoter and was purified and immobilized in a fusion form, and its reusability was investigated.ResultsThe fusion protein CBM-GAD was expressed in Escherichia coli DH5α carrying pCROCB-gadB, which contained promoter P_rpoS, cbm3 (family 3 carbohydrate-binding module from Clostridium thermocellum) coding sequence, the gadB gene from L. fermentum YS2 coding for GAD, and the T7 terminator. After a one-step purification of CBM-GAD using regenerated amorphous cellulose (RAC) as an adsorbent, SDS-PAGE analysis revealed a clear band of 71 kDa; the specific activity of the purified fusion protein CBM-GAD reached 83.6 ± 0.7 U·mg^-1. After adsorption onto RAC, the immobilized GAD with CBM3 tag was repeatedly used for GABA synthesis. The protein-binding capacity of RAC was 174 ± 8 mg·g^-1. The immobilized CBM-GAD could repeatedly catalyze GABA synthesis, and 8% of the initial activities was retained after 10 uses. We tested the conversion of monosodium glutamate to GABA by the immobilized enzyme; the yield reached 5.15 g/L and the productivity reached 3.09 g/L·h.ConclusionsRAC could be used as an adsorbent in one-step purification and immobilization of CBM-GAD, and the immobilized enzyme could be repeatedly used to catalyze the conversion of glutamate to GABA.     

Microbial transformation of ginsenosides extracted from Panax ginseng adventitious roots in an airlift bioreactor

BackgroundGinsenoside is the most important secondary metabolite in ginseng. Natural sources of wild ginseng have been overexploited. Although root culture can reduce the length of the growth cycle of ginseng, the number of species of ginsenosides is reduced and their contents are lower in the adventitious roots of ginseng than in the roots of ginseng cultivated in the field.ResultsIn this study, 147 strains of β-glucosidase-producing microorganisms were isolated from soil. Of these, strain K35 showed excellent activity for converting major ginsenosides into rare ginsenosides, and a NCBI BLAST of its 16S rDNA gene sequence showed that it was most closely related to Penicillium sp. (HQ608083.1). Strain K35 was used to ferment the adventitious root extract, and the fermentation products were analyzed by high-performance liquid chromatography. The results showed that the content of the rare ginsenoside CK was 0.253 mg mL^-1 under the optimal converting conditions of 9 d of fermentation at pH 7.0 in LL medium, which was significantly higher than that in the adventitious roots of ginseng.ConclusionThese findings may not only solve the problem of low productivity of metabolite in ginseng root culture but may also result in the development of a new valuable method of manufacturing ginsenoside CK.     

Bioaugmentation of latex rubber sheet wastewater treatment with stimulated indigenous purple nonsulfur bacteria by fermented pineapple extract

BackgroundTreating latex rubber sheet wastewater often leads to the generation of a rotten-egg odor from toxic H₂S. To increase the treatment efficiency and eliminate H₂S, purple nonsulfur bacteria (PNSB), prepared by supplementing non-sterile rubber sheet wastewater (RAW) with fermented pineapple extract (FPE), were used to treat this wastewater under microaerobic light conditions. The following 3 independent variables: chemical oxygen demand (COD), initial pH and FPE dose were investigated using the Box–Behnken design to find optimal conditions for stimulating the growth of indigenous PNSB (PNSBsi).ResultsThe addition of 2.0% FPE into RAW, which had a COD of 2000 mg L^- 1 and an initial pH of 7.0, significantly decreased oxidation reduction potential (ORP) value and stimulated PNSBsi to reach a maximum of 7.8 log cfu mL^- 1 within 2 d. Consequently, these PNSBsi, used as inoculants, were investigated for their ability to treat the wastewater under microaerobic light conditions. A central composite design was used to determine the optimal conditions for the wastewater treatment. These proved to be 7% PNSBsi, 0.8% FPE and 4 d retention time and this combination resulted in a reduction of 91% for COD, 75% for suspended solids, 61% for total sulfide while H₂S was not detected. Results of abiotic control and treatment sets indicated that H₂S was produced by heterotrophic bacteria and it was then effectively deactivated by PNSBsi.ConclusionsThe stimulation of PNSB growth by FPE under light condition was to lower ORP, and PNSBsi proved to be effective for treating the wastewater.     

Kernel size and weight affected by three plant bioregulators applied at bloom to Non Pareil and Carmel almond cultivars

BackgroundThe yield of almonds [Prunus dulcis (Mill.) D.A. Webb] could be low due to climatic problems and any factor improving kernel size and weight, such as the use of plant bioregulators (PBRs), should be beneficial.ResultsThree plant bioregulators: 24-epibrassinolide (BL), gibberellic acid (GA₃) and kinetin (KN) were applied at three spray concentrations to Non Pareil and Carmel cultivars, at two phenological stages during bloom, in the 2014 and 2015 seasons. The results showed significant differences (P < 0.0001). For total dry weight of Non Pareil, the best treatment was BL (30 mg·L^-1), with an average of 1.45 g, while the control was 1.30 g, at pink button during 2015. For Carmel, the best dry weight was 1.23 g, achieved with BL (30 mg·L^-1) at fallen petals in both seasons. The average dry weight of the controls varied between 1.13 and 1.18 g. The greatest almond lengths and widths in Non Pareil were 24.98 mm and 15.05 mm, achieved with BL (30 mg·L^-1) and KN (50 μL·L^-1) treatments, respectively, applied at pink button in 2015. In Carmel, the greatest length and width were 24.38 and 13.44 mm, obtained with BL (30 mg·L^-1) applied at the stages of pink button and fallen petals, respectively, in 2015. The control reached lengths between 22.33 and 23.38 mm, and widths between 11.99 and 12.93 mm.ConclusionsThe use of the bioregulators showed significant favorable effects on dry weight, length and width of kernels at harvest, in both cultivars.     

Biomineralization of a calcifying ureolytic bacterium Microbacterium sp. GM-1

BackgroundBiomineralization is a significant process performed by living organisms in which minerals are produced through the hardening of biological tissues. Herein, we focus on calcium carbonate precipitation, as part of biomineralization, to be used in applications for environmental protection, material technology, and other fields. A strain GM-1, Microbacterium sp. GM-1, isolated from active sludge, was investigated for its ability to produce urease and induce calcium carbonate precipitation in a metabolic process.ResultsIt was discovered that Microbacterium sp. GM-1 resisted high concentrations of urea up to 60 g/L. In order to optimize the calcification process of Microbacterium sp. GM-1, the concentrations of Ni^2 + and urea, pH value, and culture time were analyzed through orthogonal tests. The favored calcite precipitation culture conditions were as follows: the concentration of Ni^2 + and urea were 50 μM and 60 g/L, respectively, pH of 10, and culture time of 96 h. Using X-ray diffraction analysis, the calcium carbonate polymorphs produced by Microbacterium sp. GM-1 were proven to be mainly calcite.ConclusionsThe results of this research provide evidence that Microbacterium sp. GM-1 can biologically induce calcification and suggest that strain GM-1 may play a potential role in the synthesis of new biominerals and in bioremediation or biorecovery.     

Strategy of oxygen transfer coefficient control on the l-erythrulose fermentation by newly isolated Gluconobacter kondonii

BackgroundThe effect of diverse oxygen transfer coefficient on the l-erythrulose production from meso-erythritol by a newly isolated strain, Gluconobacter kondonii CGMCC8391 was investigated. In order to elucidate the effects of volumetric mass transfer coefficient (k_La) on the fermentations, baffled and unbaffled flask cultures, and fed-batch cultures were developed in present work.ResultsWith the increase of the k_La value in the fed-batch culture, l-erythrulose concentration, productivity and yield were significantly improved, while cell growth was not the best in the high k_La. Thus, a two-stage oxygen supply control strategy was proposed, aimed at achieving high concentration and high productivity of l-erythrulose. During the first 12 h, k_La was controlled at 40.28 h^-1 to obtain high value for cell growth, subsequently k_La was controlled at 86.31 h^-1 to allow for high l-erythrulose accumulation.ConclusionsUnder optimal conditions, the l-erythrulose concentration, productivity, yield and DCW reached 207.9 ± 7.78 g/L, 6.50 g/L/h, 0.94 g/g, 2.68 ± 0.17 g/L, respectively. At the end of fermentation, the l-erythrulose concentration and productivity were higher than those in the previous similar reports.     

On the fermentative behavior of auxotrophic strains of Saccharomyces cerevisiae

BackgroundThe selection of new yeast strains could lead to improvements in bioethanol production. Here, we have studied the fermentative capacity of different auxotrophic mutants of Saccharomyces cerevisiae, which are routinely used as hosts for the production of heterologous proteins. It has recently been found that these strains exhibit physiological alterations and peculiar sensitivities with respect to the parental prototrophic strains from which they derive. In this work the performance of auxotrophic S. cerevisiae CEN.PK strains was compared to the corresponding prototrophic strain, to S. cerevisiae T5bV, a strain isolated from grape must and to another auxotrophic strain, S. cerevisiae BY4741.ResultsThe results indicate that the fermentative capacity of strains grown in 2% glucose was similar in all the strains tested. However, in 15% initial glucose, the auxotrophic strains exhibited a more than doubled ethanol yield on biomass (10 g g^- 1_dw) compared to the prototrophic strains (less than 5 g g^- 1_dw). Other tests have also evidenced that in medium depletion conditions, ethanol production continues after growth arrest.ConclusionsThe results highlight the capacity of auxotrophic yeast strains to produce ethanol per mass unit, in a higher amount with respect to the prototrophic ones. This leads to potential applications for auxotrophic strains of S. cerevisiae in the production of ethanol in both homogeneous and heterogeneous phases (immobilized systems). The higher ethanol yield on biomass would be advantageous in immobilized cell systems, as a reduced yeast biomass could greatly reduce the mass transfer limitations through the immobilization matrix.     

Xylitol production and furfural consumption by a wild type Geotrichum sp.

BackgroundXylitol is a five carbons polyol with promising medical applications. It can be obtained from chemical d-xylose reduction or by microbial fermentation of Sugarcane Bagasse Hemicellulosic Hydrolysate. For this last process, some microbial inhibitors, as furfural, constitute severe bottleneck. In this case, the use of strains able to produce xylitol simultaneously to furfural neutralization is an interesting alternative. A wild-type strain of Geotrichum sp. was detected with this ability, and its performance in xylitol production and furfural consumption was evaluated. Furthermore, were analyzed its degradation products.ResultsGeotrichum sp. produced xylitol from d-xylose fermentation with a yield of 0.44 g·g^-1. Furfural was fully consumed in fermentation assay and when provided in the medium until concentration of 6 g·L^-1. The furfural degradation product is not an identified molecule, presenting a molecular weight of 161 g·mol^-1, an uncommon feature for the microbial metabolism of this product.ConclusionThis strain presents most remarkable potential in performing furfural consumption simultaneous to xylitol production. Subsequent efforts must be employed to establish bioprocess to simultaneous detoxification and xylitol production by Geotrichum sp.     

Utilization of a waste glycerol fraction using and reusing immobilized Gluconobacter oxydans ATCC 621 cell extract

BackgroundDepletion of petroleum resources has enforced the search for alternative sources of renewable energy. Introduction of biofuels into the market was expected to become a solution to this disadvantageous situation. Attempts to cover fuel demand have, however, caused another severe problem—the waste glycerol generated during biodiesel production at a concentration of approximately 10% w/w. This, in turn, prompted a global search for effective methods of valorization of the waste fraction of glycerol.ResultsUtilization of the waste fraction at 48 h with an initial glycerol concentration of 30 g·L^-1 and proceeding with 62% efficiency enabled the production of 9 g·L^-1 dihydroxyacetone at 50% substrate consumption. The re-use of the immobilized biocatalyst resulted in a similar concentration of dihydroxyacetone (8.7 g·L^-1) in two-fold shorter time, with an efficiency of 85% and lower substrate consumption (35%).ConclusionsThe method proposed in this work is based on the conversion of waste glycerol to dihydroxyacetone in a reaction catalyzed by immobilized Gluconobacter oxydans cell extract with glycerol dehydrogenase activity, and it could be an effective way to convert waste glycerol into a valuable product.     

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

BackgroundBiohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated.ResultsThe methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15–20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P < 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P < 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield.ConclusionPreventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.     

1-Deoxynojirimycin from Bacillus subtilis improves antioxidant and antibacterial activities of juvenile Yoshitomi tilapia

BackgroundJuvenile Yoshitomi tilapia is often infected by pathogens and results in low-level survival rate. Bacillus subtilis, as a probiotic, may have beneficial effects on Y. tilapia with compound 1-deoxynojirimycin (DNJ), which has antibacterial activities. The effects of dietary probiotic supplementation on Y. tilapias were evaluated.ResultsJuvenile Y. tilapia was fed with B. subtilis for 56 d. Y. tilapia was infected by Aeromonas hydrophila and survival rate was compared. Dietary B. subtilis increased weight gain rate, specific growth, food conversion ratios and food intake rate of Y. tilapia. The diet improved the cumulative survival rate (CSR) of juvenile Y. tilapia when the concentration of B. subtilis was more than 2.05 × 10¹⁰ cfu/kg and CSR reached a maximum rate when the concentration of bacillus was 4.23 × 10¹⁰ (P < 0.05). Meanwhile, B. subtilis improved total antioxidant capacity (TAC), spleen index, the activities of serum lysozyme, alkaline phosphatase (ALP), superoxide dismutase (SOD) and catalase (CAT) (P < 0.05). In contrast, B. subtilis reduced serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA) and C3 complement (P < 0.05). DNJ was isolated from secondary metabolisms and proved to increase the levels of SOD, CAT and reduce the levels of AST, ALT and MDA at cell levels. After A. hydrophila infection, DNJ prevented the reduction in survival rate of Y. tilapia (P < 0.05).Conclusions1-Deoxynojirimycin from Bacillus subtilis can be used to improve the growth performance of juvenile Y. tilapia by affecting its antioxidant and antibacterial activities.     

Poly(dl-lactide)-degrading enzyme production by immobilized Actinomadura keratinilytica strain T16-1 in a 5-L fermenter under various fermentation processes

BackgroundPoly(dl-lactic acid), or PDLLA, is a biodegradable polymer that can be hydrolyzed by various types of enzymes. The protease produced by Actinomadura keratinilytica strain T16-1 was previously reported to have PDLLA depolymerase activity. However, few studies have reported on PDLLA-degrading enzyme production by bacteria. Therefore, the aims of this study were to determine a suitable immobilization material for PDLLA-degrading enzyme production and optimize PDLLA-degrading enzyme production by using immobilized A. keratinilytica strain T16-1 under various fermentation process conditions in a stirrer fermenter.ResultsAmong the tested immobilization materials, a scrub pad was the best immobilizer, giving an enzyme activity of 30.03 U/mL in a shake-flask scale. The maximum enzyme activity was obtained at aeration 0.25 vvm, agitation 170 rpm, 45°C, and 48 h of cultivation time. Under these conditions, a PDLLA-degrading enzyme production of 766.33 U/mL with 15.97 U/mL·h productivity was observed using batch fermentation in a 5-L stirrer fermenter. Increased enzyme activity and productivity were observed in repeated-batch (942.67 U/mL and 19.64 U/mL·h) and continuous fermentation (796.43 U/mL and 16.58 U/mL·h) at a dilution rate of 0.013/h. Scaled-up production of the enzyme in a 10-L stirrer bioreactor using the optimized conditions showed a maximum enzyme activity of 578.67 U/mL and a productivity of 12.06 U/mL·h.ConclusionsThis research successfully scaled-up the enzyme production to 5 and 10 L in a stirrer fermenter and is helpful for many applications of poly(lactic acid).     

Bioconversion of mixed free fatty acids to poly-3-hydroxyalkanoates by Pseudomonas putida BET001 and modeling of its fermentation in shake flasks

BackgroundThe paper reports on the utilization of palm kernel oil (PKO) as a low cost renewable substrate for medium-chain-length poly-3-hydroxyalkanoates (mcl-PHA) production by Pseudomonas putida BET001. Investigation on the effects of selected key variables on growth, mixed free fatty acids consumption and mcl-PHA production by the bacterial culture in the shaken flask system were carried out along with its kinetic modeling.ResultsThe biomass production, fatty acids consumption and mcl-PHA production were found favorable when the strain was cultured in mineral medium at pH 6–7, 28°C, aeration surface-to-volume ratio of 0.4 × 10⁶ m^- 1, 250 rpm agitation rate for 48 h. Mcl-PHA production by this strain showed mixed growth and non-growth associated components as described by Luedeking–Piret kinetic model.ConclusionThe findings of this study provided add to the literature on key variables in for achieving good microbial growth and mcl-PHA production in shake flasks culture. In addition, suitable kinetic model to describe cultivation in this system was also presented.     

One-pot synthesis of graphene–chitosan nanocomposite modified carbon paste electrode for selective determination of dopamine

BackgroundA simple, rapid, low-cost and environmentally friendly method was developed to determine dopamine (DA) in the presence of ascorbic (AA) and uric acid (UA) based on a novel technique to prepare a graphene–chitosan (GR–CS) nanocomposite and modify it on the surface of carbon paste electrode (CPE). For our design, CS acts as a media to disperse and stabilize GR, and then GR plays a key role to selective and sensitive determination of DA.ResultsUnder physiological conditions, the linear range for dopamine was determined from 1 × 10^- 4 to 2 × 10^- 7 mol/L with a good correlation coefficient of 0.9961 in the presence of 1000-fold interference of AA and UA. The detection limit was estimated to be 9.82 × 10^- 8 mol/L (S/N = 3). In order to study the stability and reproducibility, GR/CS/CPE underwent successive measurements in 10 times and then tested once a d for 30 d. The result exhibited 98.25% and 91.62% activities compared with the original peak current after 10-time measurements and 30-d storage.ConclusionThe GR/CS/CPE has wide linear concentration range, low detection limit, and good reproducibility and stability, which suggests that our investigations provide a promising alternative for clinic DA determination.     

A quantitative PCR approach for determining the ribosomal DNA copy number in the genome of Agave tequila Weber

BackgroundAgave tequilana has a great economic importance in Mexico in order to produce alcoholic beverages and bioenergy. However, in this species the structure and organization of the rDNAs in the genome are limited, and it represents an obstacle both in their genetic research and improvement as well. rDNA copy number variations per eukaryotic genome have been considered as a source of genetic rearrangements. In this study, the copy number of 18S and 5S rDNAs in the A. tequilana genome was estimated, and an absolute quantitative qPCR assay and genome size was used. In addition, an association between the rDNAs copy number and physical mapping was performed to confirm our results.ResultsThe analysis were successfully applied to determine copy number of 18S and 5S rDNAs in A. tequilana genome, showing high reproducibility with coefficient of variation (CV) values of 0.014–0.0129%, respectively. A variation of 51 times in the copy number the 18s regarding 5s rDNA was found, thus contributing to genome size of 1.47 and 8.38 × 10^-3%, respectively. Similarly, data show a linear relationship (R [2] = 0.992) between rDNA copy number and the detected signals for each of the loci by FISH. The comparison of the rDNA copy number of agave showed differential relationship with other organisms and it may be due to evolutionary ecology.ConclusionsResults show that the proposed method a) can correctly detect the rDNA copy number, b) could be used as species-specific markers and c) might help in understanding the genetic diversity, genome organization and evolution of this species.     

Characterization of OAZ1 and its potential functions in goose follicular development

BackgroundOrnithine decarboxylase antizyme 1 (OAZ1) is an important regulator of polyamine synthesis and uptake. Our previous studies indicated that high OAZ1 expression in the ovaries of laying geese is responsible for poor egg production. In the present study, the molecular characterization of goose OAZ1 gene was analyzed, as well as the expression profile in various follicular tissues.ResultsAn 873-bp cDNA sequence of the OAZ1 gene (Accession No. KC845302) with a + 1 frameshift site (+ 175T) was obtained. The sequence consisted of a 652-bp two overlapping open reading frames (a putative protein with 216 amino acids). The OAZ domain, OAZ signature and OAZ super family domain were prominent conserved regions among species. As the follicle size increased, OAZ1 abundance showed an increasing trend during follicular development, while it decreased during follicular regression. The level of OAZ1 mRNA expression was the lowest in the fifth largest preovulatory follicle, and was 0.65-fold compared to the small white follicle (P < 0.05). OAZ1 mRNA expression in the largest preovulatory and postovulatory follicle was 2.11- and 2.49-fold compared to the small white follicle, respectively (P < 0.05).ConclusionsThe goose OAZ1 structure confirms that OAZ1 plays an important role in ornithine decarboxylase-mediated regulation of polyamine homeostasis. Our findings provide an evidence for a potential function of OAZ1 in follicular development, ovulation and regression.     

Improvement of hydrogen yield of ethanol-producing Escherichia coli recombinants in acidic conditions

BackgroundAn effective single culture with high glycerol consumption and hydrogen and ethanol coproduction yield is still in demand. A locally isolated glycerol-consuming Escherichia coli SS1 was found to produce lower hydrogen levels under optimized ethanol production conditions. Molecular approach was proposed to improve the hydrogen yield of E. coli SS1 while maintaining the ethanol yield, particularly in acidic conditions. Therefore, the effect of an additional copy of the native hydrogenase gene hycE and recombinant clostridial hydrogenase gene hydA on hydrogen production by E. coli SS1 at low pH was investigated.ResultsRecombinant E. coli with an additional copy of hycE or clostridial hydA was used for fermentation using 10 g/L (108.7 mmol/L) of glycerol with an initial pH of 5.8. The recombinant E. coli with hycE and recombinant E. coli with hydA showed 41% and 20% higher hydrogen yield than wild-type SS1 (0.46 ± 0.01 mol/mol glycerol), respectively. The ethanol yield of recombinant E. coli with hycE (0.50 ± 0.02 mol/mol glycerol) was approximately 30% lower than that of wild-type SS1, whereas the ethanol yield of recombinant E. coli with hydA (0.68 ± 0.09 mol/mol glycerol) was comparable to that of wild-type SS1.ConclusionsInsertion of either hycE or hydA can improve the hydrogen yield with an initial pH of 5.8. The recombinant E. coli with hydA could retain ethanol yield despite high hydrogen production, suggesting that clostridial hydA has an advantage over the hycE gene in hydrogen and ethanol coproduction under acidic conditions. This study could serve as a useful guidance for the future development of an effective strain coproducing hydrogen and ethanol.     

Statistical optimization of thermo-alkali stable xylanase production from Bacillus tequilensis strain ARMATI

BackgroundXylanase from bacteria finds use in prebleaching process and bioconversion of lignocelluloses into feedstocks. The xylanolytic enzyme brings about the hydrolysis of complex biomolecules into simple monomer units. This study aims to optimize the cellulase-free xylanase production and cell biomass of Bacillus tequilensis strain ARMATI using response surface methodology (RSM).ResultsStatistical screening of medium constituents and the physical factors affecting xylanase and biomass yield of the isolate were optimized by RSM using central composite design at N = 30, namely 30 experimental runs with 4 independent variables. The central composite design showed 3.7 fold and 1.5 fold increased xylanase production and biomass yield of the isolate respectively compared to ‘one factor at a time approach’, in the presence of the basal medium containing birchwood xylan (1.5% w/v) and yeast extract (1% w/v), incubated at 40°C for 24 h. Analysis of variance (ANOVA) revealed high coefficient of determination (R²) of 0.9978 and 0.9906 for the respective responses at significant level (p < 0.05). The crude xylanase obtained from the isolate showed stability at high temperature (60°C) and alkaline condition (pH 9) up to 4 h of incubation.ConclusionsThe cellulase-free xylanase showed an alkali-tolerant and thermo-stable property with potentially applicable nature at industrial scale. This statistical approach established a major contribution in enzyme production from the isolate by optimizing independent factors and represents a first reference on the enhanced production of thermo-alkali stable cellulase-free xylanase from B. tequilensis.