Myco-synthesized copper oxide nanoparticles using harnessing metabolites of endophytic fungal strain Aspergillus terreus: an insight into antibacterial, anti-Candida, biocompatibility, anticancer, and antioxidant activities.

BACKGROUND: The overuse of antibiotics leads to the emergence of antibiotic-resistant microbes which causes high mortality worldwide. Therefore, the synthesis of new active compounds has multifunctional activities are the main challenge. Nanotechnology provides a solution for this issue. METHOD: The endophytic fungal strain Aspergillus terreus BR.1 was isolated from the healthy root of Allium sativum and identified using internal transcribed spacer (ITS) sequence analysis. The copper oxide nanoparticles (CuO-NPs) were synthesized by harnessing the metabolites of the endophytic fungal strain. The UV-Visble spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), Transmission electron micrscopy (TEM), Energy dispersive X-ray (EDX), X-ray diffraction (XRD), Dynamic light scattering (DLS), and zeta potential (ζ) were used for the characterization of synthesized CuO-NPs. The activity against different pathogenic bacteria and Candida species were investigated by agar well-diffusion method. The biocombatibility and anticancer activity were assessed by MTT assay method. The scavenging of DPPH was used to investigate the antioxidant activity of synthesized CuO-NPs. RESULTS: Data showed the successful formation of crystalline nature and spherical shape CuO-NPs with sizes in the ranges of 15-55 nm. The EDX reveals that the as-formed sample contains ions of C, O, Cl, and Cu with weight percentages of 18.7, 23.82, 11.31, and 46.17%, respectively. The DLS and ζ-potential showed high homogeneity and high stability of synthesized CuO-NPs with a polydispersity index (PDI) of 0.362 and ζ-value of - 26.6 mV. The synthesized CuO-NPs exhibited promising antibacterial and anti-Candida activity (concentration-dependent) with minimum inhibitory concentration (MIC) values in the ranges of 25-50 µg mL-1. Moreover, the fungal mediated-CuO-NPs targeted cancer cells of MCF7 and PC3 at low IC50 concentrations of 159.2 ± 4.5 and 116.2 ± 3.6 µg mL-1, respectively as compared to normal cells (Vero and Wi38 with IC50 value of 220.6 ± 3.7 and 229.5 ± 2.1 µg mL-1, respectively). The biosynthesized CuO-NPs showed antioxidant activity as detected by the DPPH method with scavenging percentages of 80.5 ± 1.2% at a concentration of 1000 µg mL-1 and decreased to 20.4 ± 4.2% at 1.9 µg mL-1 as compared to ascorbic acid (control) with scavenging activity of 97.3 ± 0.2 and 37.5 ± 1.3% at the same concentrations, respectively. CONCLUSION: The fungal mediated-CuO-NPs exhibited promising activity and can be integrated into various biomedical and theraputic applications.

Antimicrobial, Antiviral, and In-Vitro Cytotoxicity and Mosquitocidal Activities of Portulaca oleracea-Based Green Synthesis of Selenium Nanoparticles.

The aqueous extract of Portulaca oleracea was used as a biocatalyst for the reduction of Na2SeO3 to form Se-NPs that appeared red in color and showed maximum surface plasmon resonance at a wavelength of 266 nm, indicating the successful Phyto-fabrication of Se-NPs. A FT-IR chart clarified the role of plant metabolites such as proteins, carbohydrates, and amino acids in capping and stabilizing Se-NPs. TEM, SAED, and XRD analyses indicated the formation of spherical, well-arranged, and crystalline Se-NPs with sizes in the range of 2-22 nm. SEM-EDX mapping showed the maximum peaks of Se at 1.4, 11.3, and 12.4 KeV, with weight and atomic percentages of 36.49 and 30.39%, respectively. A zeta potential of -43.8 mV also indicated the high stability of the synthesized Se-NPs. The Phyto-synthesized Se-NPs showed varied biological activities in a dose-dependent manner, including promising activity against pathogenic bacteria and Candida species with varied MIC values in the range of 12.5-50 µg·mL-1. Moreover, the Se-NPs showed antiviral activity toward HAV and Cox-B4, with percentages of 70.26 and 62.58%, respectively. Interestingly, Se-NPs showed a target orientation to cancer cell lines (HepG2) with low IC50 concentration at 70.79 ± 2.2 µg·mL-1 compared to normal cell lines (WI-38) with IC50 at165.5 ± 5.4 µg·mL-1. Moreover, the as-formed Se-NPs showed high activity against various instar larvae I, II, III, and IV of Culex pipiens, with the highest mortality percentages of 89 ± 3.1, 73 ± 1.2, 68 ± 1.4, and 59 ± 1.0%, respectively, at 50 mg L-1. Thus, P. oleracea-based Se-NPs would be strong potential antimicrobial, anti-viral, anti-cancer, and anti-insect agents in the pharmaceutical and biomedical industries.

Light enhanced the antimicrobial, anticancer, and catalytic activities of selenium nanoparticles fabricated by endophytic fungal strain, Penicillium crustosum EP-1.

Selenium nanoparticles (Se-NPs) has recently received great attention over owing to their superior optical properties and wide biological and biomedical applications. Herein, crystallographic and dispersed spherical Se-NPs were green synthesized using endophytic fungal strain, Penicillium crustosum EP-1. The antimicrobial, anticancer, and catalytic activities of biosynthesized Se-NPs were investigated under dark and light (using Halogen tungsten lamp, 100 Watt, λ > 420 nm, and light intensity of 2.87 W m-2) conditions. The effect of Se-NPs was dose dependent and higher activities against Gram-positive and Gram-negative bacteria as well different Candida spp. were attained in the presence of light than obtained under dark conditions. Moreover, the viabilities of two cancer cells (T47D and HepG2) were highly decreased from 95.8 ± 2.9% and 93.4 ± 3.2% in dark than those of 84.8 ± 2.9% and 46.4 ± 3.3% under light-irradiation conditions, respectively. Significant decreases in IC50 values of Se-NPs against T47D and HepG2 were obtained at 109.1 ± 3.8 and 70.4 ± 2.5 µg mL-1, respectively in dark conditions than 19.7 ± 7.2 and 4.8 ± 4.2 µg mL-1, respectively after exposure to light-irradiation. The photoluminescence activity of Se-NPs revealed methylene blue degradation efficiency of 89.1 ± 2.1% after 210 min under UV-irradiation compared to 59.7 ± 0.2% and 68.1 ± 1.03% in dark and light conditions, respectively. Moreover, superior stability and efficient MB degradation efficiency were successfully achieved for at least five cycles.

Enhanced Antimicrobial, Cytotoxicity, Larvicidal, and Repellence Activities of Brown Algae, Cystoseira crinita-Mediated Green Synthesis of Magnesium Oxide Nanoparticles.

Herein, the metabolites secreted by brown algae, Cystoseira crinita, were used as biocatalyst for green synthesis of magnesium oxide nanoparticles (MgO-NPs). The fabricated MgO-NPs were characterized using UV-vis spectroscopy, Fourier transforms infrared spectroscopy (FT-IR), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy linked with energy-dispersive X-ray (SEM-EDX), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). Data showed successful formation of crystallographic and spherical MgO-NPs with sizes of 3-18 nm at a maximum surface plasmon resonance of 320 nm. Moreover, EDX analysis confirms the presence of Mg and O in the sample with weight percentages of 54.1% and 20.6%, respectively. Phyco-fabricated MgO-NPs showed promising activities against Gram-positive bacteria, Gram-negative bacteria, and Candida albicans with MIC values ranging between 12.5 and 50 µg mL-1. The IC50 value of MgO-NPs against cancer cell lines (Caco-2) was 113.4 µg mL-1, whereas it was 141.2 µg mL-1 for normal cell lines (Vero cell). Interestingly, the green synthesized MgO-NPs exhibited significant larvicidal and pupicidal activity against Musca domestica. At 10 µg mL-1 MgO-NPs, the highest mortality percentages were 99.0%, 95.0%, 92.2%, and 81.0% for I, II, III instars' larvae, and pupa of M. domestica, respectively, with LC50 values (3.08, 3.49, and 4.46 µg mL-1), and LC90 values (7.46, 8.89, and 10.43 µg mL-1), respectively. Also, MgO-NPs showed repellence activity for adults of M. domestica at 10 µg mL-1 with 63.0%, 77.9%, 84.9%, and 96.8% after 12, 24, 48, and 72 h, respectively.

Harnessing Bacterial Endophytes for Promotion of Plant Growth and Biotechnological Applications: An Overview.

Endophytic bacteria colonize plants and live inside them for part of or throughout their life without causing any harm or disease to their hosts. The symbiotic relationship improves the physiology, fitness, and metabolite profile of the plants, while the plants provide food and shelter for the bacteria. The bacteria-induced alterations of the plants offer many possibilities for biotechnological, medicinal, and agricultural applications. The endophytes promote plant growth and fitness through the production of phytohormones or biofertilizers, or by alleviating abiotic and biotic stress tolerance. Strengthening of the plant immune system and suppression of disease are associated with the production of novel antibiotics, secondary metabolites, siderophores, and fertilizers such as nitrogenous or other industrially interesting chemical compounds. Endophytic bacteria can be used for phytoremediation of environmental pollutants or the control of fungal diseases by the production of lytic enzymes such as chitinases and cellulases, and their huge host range allows a broad spectrum of applications to agriculturally and pharmaceutically interesting plant species. More recently, endophytic bacteria have also been used to produce nanoparticles for medical and industrial applications. This review highlights the biotechnological possibilities for bacterial endophyte applications and proposes future goals for their application.

Distribution of Extended-Spectrum ß-Lactamase (ESBL)-Encoding Genes among Multidrug-Resistant Gram-Negative Pathogens Collected from Three Different Countries.

The incidence of Extended-spectrum ß-lactamase (ESBL)-encoding genes (blaCTX-M and blaTEM) among Gram-negative multidrug-resistant pathogens collected from three different countries was investigated. Two hundred and ninety-two clinical isolates were collected from Egypt (n = 90), Saudi Arabia (n = 162), and Sudan (n = 40). Based on the antimicrobial sensitivity against 20 antimicrobial agents from 11 antibiotic classes, the most resistant strains were selected and identified using the Vitek2 system and 16S rRNA gene sequence analysis. A total of 85.6% of the isolates were found to be resistant to more than three antibiotic classes. The ratios of the multidrug-resistant strains for Egypt, Saudi Arabia, and Sudan were 74.4%, 90.1%, and 97.5%, respectively. Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa showed inconstant resistance levels to the different classes of antibiotics. Escherichia coli and Klebsiella pneumoniae had the highest levels of resistance against macrolides followed by penicillins and cephalosporin, while Pseudomonas aeruginosa was most resistant to penicillins followed by classes that varied among different countries. The isolates were positive for the presence of the blaCTX-M and blaTEM genes. The blaCTX-M gene was the predominant gene in all isolates (100%), while blaTEM was detected in 66.7% of the selected isolates. This work highlights the detection of multidrug-resistant bacteria and resistant genes among different countries. We suggest that the medical authorities urgently implement antimicrobial surveillance plans and infection control policies for early detection and effective prevention of the rapid spread of these pathogens.

Developing Liquid Rhizobium Inoculants with Enhanced Long-Term Survival, Storage Stability, and Plant Growth Promotion Using Ectoine Additive.

Liquid microbial inoculants have recently received great attention due to their vital roles for sustainable agricultural practices. However, long-term conservation under ambient temperature conditions and deleterious environmental factors might negatively impact microbial cell survival and limit their efficacy in the field. Thus, developing efficient liquid formulation providing prolonged survival of rhizobia in the final product and after an application is crucial. Therefore, this study investigates the effect of various additives on the long-term survival of rhizobia stored in liquid cultures at room temperature (25 °C) for 12 months. Various yeast sucrose media amended with polyvinylpyrrolidone (PVP) or gum arabic as colloidal agents in combination with ectoine (as a compatible solute) and/or glycerol were evaluated. A dramatic decline in viable cell count was obtained in formulas amended only with PVP from Log 8.5 to Log 5 in the first six months and then to Log 1.5 after 12 months. In contrast, rhizobia stored at PVP-based formulas amended with 10 mg L‒1 ectoine exhibited almost constant survival level till the end of the storage period. The same trend was obtained using formulas based on gum arabic as a colloidal dispersing agent; however, less decline in cell count using a formula containing gum arabic alone as compared to using PVP. On the other hand, PVP based formulas exhibited higher viscosity compared with another formula. Increased viscosity till the 8th month of storage was achieved in the presence of ectoine indicating the increase of exopolymeric substances production. Electrophoretic protein pattern of rhizobial cells (stored for 12 months) exhibited several low molecular weight protein bands in cells stored in PVP based formula with ectoine as compared to the other treatments. Thus, the amendment of the liquid formulation of rhizobia bioinoculant with PVP plus ectoine not only improved cell survival but also enhanced the culture viscosity and consequently ameliorate the colonization and performance of rhizobial inoculants.

Transcriptome profile of carbon catabolite repression in an efficient l-(+)-lactic acid-producing bacterium Enterococcus mundtii QU25 grown in media with combinations of cellobiose, xylose, and glucose.

Enterococcus mundtii QU25, a non-dairy lactic acid bacterium of the phylum Firmicutes, is capable of simultaneously fermenting cellobiose and xylose, and is described as a promising strain for the industrial production of optically pure l-lactic acid (≥ 99.9%) via homo-fermentation of lignocellulosic hydrolysates. Generally, Firmicutes bacteria show preferential consumption of sugar (usually glucose), termed carbon catabolite repression (CCR), while hampering the catabolism of other sugars. In our previous study, QU25 exhibited apparent CCR in a glucose-xylose mixture phenotypically, and transcriptional repression of the xylose operon encoding initial xylose metabolism genes, likely occurred in a CcpA-dependent manner. QU25 did not exhibit CCR phenotypically in a cellobiose-xylose mixture. The aim of the current study is to elucidate the transcriptional change associated with the simultaneous utilization of cellobiose and xylose. To this end, we performed RNA-seq analysis in the exponential growth phase of E. mundtii QU25 cells grown in glucose, cellobiose, and/or xylose as either sole or co-carbon sources. Our transcriptomic data showed that the xylose operon was weakly repressed in cells grown in a cellobiose-xylose mixture compared with that in cells grown in a glucose-xylose mixture. Furthermore, the gene expression of talC, the sole gene encoding transaldolase, is expected to be repressed by CcpA-mediated CCR. QU25 metabolized xylose without using transaldolase, which is necessary for homolactic fermentation from pentoses using the pentose-phosphate pathway. Hence, the metabolism of xylose in the presence of cellobiose by QU25 may have been due to 1) sufficient amounts of proteins encoded by the xylose operon genes for xylose metabolism despite of the slight repression of the operon, and 2) bypassing of the pentose-phosphate pathway without the TalC activity. Accordingly, we have determined the targets of genetic modification in QU25 to metabolize cellobiose, xylose and glucose simultaneously for application of the lactic fermentation from lignocellulosic hydrolysates.

Correction to: Dynamic simulation of continuous mixed sugar fermentation with increasing cell retention time for lactic acid production using Enterococcus mundtii QU 25.

[This corrects the article DOI: 10.1186/s13068-020-01752-6.].

Dynamic simulation of continuous mixed sugar fermentation with increasing cell retention time for lactic acid production using Enterococcus mundtii QU 25.

BACKGROUND: The simultaneous and effective conversion of both pentose and hexose in fermentation is a critical and challenging task toward the lignocellulosic economy. This study aims to investigate the feasibility of an innovative co-fermentation process featuring with a cell recycling unit (CF/CR) for mixed sugar utilization. A l-lactic acid-producing strain Enterococcus mundtii QU 25 was applied in the continuous fermentation process, and the mixed sugars were utilized at different productivities after the flowing conditions were changed. A mathematical model was constructed with the experiments to optimize the biological process and clarify the cell metabolism through kinetics analysis. The structured model, kinetic parameters, and achievement of the fermentation strategy shall provide new insights toward whole sugar fermentation via real-time monitoring for process control and optimization. RESULTS: Significant carbon catabolite repression in co-fermentation using a glucose/xylose mixture was overcome by replacing glucose with cellobiose, and the ratio of consumed pentose to consumed hexose increased significantly from 0.096 to 0.461 by mass. An outstanding product concentration of 65.2 g L-1 and productivity of 13.03 g L-1 h-1 were achieved with 50 g L-1 cellobiose and 30 g L-1 xylose at an optimized dilution rate of 0.2 h-1, and the cell retention time gradually increased. Among the total lactic acid production, xylose contributed to more than 34% of the mixed sugars, which was close to the related contents in agricultural residuals. The model successfully simulated the transition of sugar consumption, cell growth, and lactic acid production among the batch, continuous process, and CF/CR systems. CONCLUSION: Cell retention time played a critical role in balancing pentose and hexose consumption, cell decay, and lactic acid production in the CF/CR process. With increasing cell concentration, consumption of mixed sugars increased with the productivity of the final product; hence, the impact of substrate inhibition was reduced. With the validated parameters, the model showed the highest accuracy simulating the CF/CR process, and significantly longer cell retention times compared to hydraulic retention time were tested.

Multifunctional properties of spherical silver nanoparticles fabricated by different microbial taxa.

This study addresses the impacts of metabolites from different microbial taxa on the fabrication and multifunctional biological properties of spherical silver nanoparticles (Ag-NPs). Three microbial taxa, a bacterial (Bacillus cereus A1-5), actinomycetes (Streptomyces noursei H1-1), and fungal (Rhizopus stolonifer A6-2) strains were used for Ag-NPs biosynthesis, whereas Streptomyces noursei is demonstrated for the first time. These isolates were identified using either 16S rRNA or ITS gene sequencing. Characterization of Ag-NPs was done using color change analysis, Uv-Vis spectroscopy, FT-IR spectroscopy, XRD, TEM, SEM-EDX, DLS, and Zeta potential analysis. All biosynthesized NPs exhibited spherical shape with different sizes ranged from 6‒50 nm, 6-30 nm and 6-40 nm for NPs obtained by A1-5, H1-1 and A6-2, respectively. The crystalline center cubic face of Ag-NPs was confirmed using XRD at 2θ values 38.08o, 44.27o, 64.41o and 77.36o. FT-IR analysis revealed varied intense absorption peaks for biomolecules required for NPs synthesize by each microbial strain. The stability of spherical Ag-NPs was confirmed due to highly DLS negative surface charge of ‒17.5mV, ‒18.9mV, and ‒15.6mV for NPs synthesized by strains A1-5, H1-1, and A6-2, respectively. Ag-NPs exhibited a broadspectrum of antibacterial activity against Gram-positive and Gram-negative bacteria with varied effectiveness. They also exhibited a cytotoxic effect against cancer cell line (caco-2) in a dose-dependent pattern with IC50 of 8.9 ± 0.5, 5.6 ± 3.0, 11.2 ± 0.5 µg/ml for NPs synthesized by strains A1-5, H1-1, and A6-2, respectively. Moreover, these spherical Ag-NPs showed larvicidal activity against the 3rd instar larvae of the dengue vector Aedes aegypti.

Pomegranate peel and moringa-based diets enhanced biochemical and immune parameters of Nile tilapia against bacterial infection by Aeromonas hydrophila.

We previously reported that Aeromonas hydrophila exhibited the highest prevalence rate amongst 182 bacterial strains isolated from naturally diseased Nile tilapia (Oreochromis niloticus) collected from El-Abassa Fish Farm, Egypt (Hassan et al., Egypt. J. Aquac., 10, 23-43, 2020). The overuse of antibiotics for controlling diseases has led to acquired antibiotics resistance of aquatic bacteria, besides the developments of human, aquatic animal and environmental risks arising from residual antibiotics. Therefore, the evaluation of safe alternative phytotherapies is of great importance. This study was conducted to evaluate and compare growth performance and immune potentiating activities of moringa (Moringa oleifera) leaves extract (Moringa LE) and pomegranate (Punica granatum) peel extract (Pomegranate PE) on Nile tilapia against infection with a pathogenic bacterium, Aeromonas hydrophila. A total of 150 Oreochromis niloticus were randomly divided into 5 groups to be fed at 3% of body weight with isonitrogenous/isoenergetic diets supplemented with Moringa LE at 0.15 and 0.25% kg-1 or Pomegranate PE at of 0.3 and 0.5% kg-1, separately. Growth performance was significantly affected by Moringa LE as compared with the control group without supplementation of plant extract, while Pomegranate PE levels did not affect growth performance. Maximum average daily gains, specific growth rate, albumin, globulin, total protein, A/G ratio, alanine amino transaminase (ALT), aspartate amino transaminase (AST), cholesterol, triglyceride, creatinine, urea, and lysozyme were analyzed. Antioxidant enzymes of catalase and superoxide dismutase were also evaluated in liver tissues. After feeding experiment, the results indicated that the addition of Moringa LE and Pomegranate PE improved lipid profile, liver and kidney functions, immune response towards the emerging bacterial diseases. Besides this, feeding the fishes on diets supplemented with Moringa LE at concentration 0.25% kg-1 showed the best growth performance, and improved immunity. Moreover, it exhibited the highest protection against bacterial infection with Aeromonas hydrophila achieving the lowest mortality rate of 10% as compared to 80% of mortality rate at the control group.

Non-carbon loss long-term continuous lactic acid production from mixed sugars using thermophilic Enterococcus faecium QU 50.

In this study, a non-sterile (open) continuous fermentation (OCF) process with no-carbon loss was developed to improve lactic acid (LA) productivity and operational stability from the co-utilization of lignocellulose-derived sugars by thermophilic Enterococcus faecium QU 50. The effects of different sugar mixtures on LA production were firstly investigated in conventional OCF at 50°C, pH 6.5 and a dilution rate of 0.20 hr-1 . The xylose consumption ratio was greatly lower than that of glucose in fermentations with glucose/xylose mixtures, indicating apparent carbon catabolite repression (CCR). However, CCR could be efficiently eliminated by feeding solutions containing the cellobiose/xylose mixture. In OCF at a dilution rate ca. 0.10 hr-1 , strain QU 50 produced 42.6 g L-1 of l-LA with a yield of 0.912 g g-1 -consumed sugars, LA yield of 0.655 g g-1 based on mixed sugar-loaded, and a productivity of 4.31 g L-1 hr-1 from simulated energy cane hydrolyzate. In OCF with high cell density by cell recycling, simultaneous and complete co-utilization of sugars was achieved with stable LA production at 60.1 ± 3.25 g L-1 with LA yield of 0.944 g g-1 -consumed sugar and LA productivity of 6.49 ± 0.357 g L-1 hr-1 . Besides this, a dramatic increase in LA yield of 0.927 g g-1 based on mixed sugar-loaded with prolonged operational stability for at least 500 hr (>20 days) was established. This robust system demonstrates an initial green step with a no-carbon loss under energy-saving toward the feasibility of sustainable LA production from lignocellulosic sugars.

Complete Genome Sequence of Enterococcus faecium QU50, a Thermophilic Lactic Acid Bacterium Capable of Metabolizing Xylose.

Herein, we report the complete genome sequence of Enterococcus faecium QU50, isolated from Egyptian soil and exhibiting intermediate susceptibility to vancomycin. The genome contains a 2,535,796-bp circular chromosome and two plasmids of 196,595 bp and 17,267 bp. IS1062-like sequences were not found.

High Improvement in Lactic Acid Productivity by New Alkaliphilic Bacterium Using Repeated Batch Fermentation Integrated with Increased Substrate Concentration.

Optically pure lactic acid (LA) is an important chemical platform that has a wide range of industrial and biotechnological applications. Improved parameters for cost effective LA production are of great interest for industrial developments. In the present study, an alkaliphilic lactic acid bacterium, BoM 1-2, was selected among 369 newly obtained bacterial isolates. It was characterized using API 50 CHL kit and identified as Enterococcus hirae BoM 1-2 by 16S rRNA gene sequence analysis. Efficient polymer-grade L-lactic acid production was achieved at pH 9.0 and 40°C. In batch fermentation strategy using 20 g L-1 glucose, 19.6 g L-1 lactic acid was obtained with volumetric productivity of 2.18 g L-1 h-1. While using 100 g L-1 glucose, 96.0 g L-1 lactic acid was obtained with volumetric productivity of 1.07 g L-1 h-1. The highest lactic acid concentration of 180.6 g L-1 was achieved in multipulse fed batch strategy with volumetric productivity of 0.65 g L-1 h-1. To achieve higher productivity, repeated fermentation processes were applied using the two different strategies. In the first strategy, the lactic acid productivity was increased from 1.97 g L-1 h-1 to 4.48 g L-1 h-1 when the total of 10 repeated runs were carried out using 60 g L-1 glucose, but lactic acid productivity decreased to 2.95 g L-1 h-1 using 100 g L-1 glucose. In second strategy, repeated fermentation coupled with gradual increase in glucose concentration from 40 to 100 g L-1 was conducted for 24 runs. A dramatic increase in LA productivity up to 39.9 g L-1 h-1 (18-fold compared to first run) was achieved using 40 g L-1 glucose while volumetric productivity ranging between 24.8 and 29.9 g L-1 h-1 was achieved using 60-100 g L-1 glucose.

Opportunities to overcome the current limitations and challenges for efficient microbial production of optically pure lactic acid.

There has been growing interest in the microbial production of optically pure lactic acid due to the increased demand for lactic acid-derived environmentally friendly products, for example biodegradable plastic (poly-lactic acid), as an alternative to petroleum-derived materials. To maximize the market uptake of these products, their cost should be competitive and this could be achieved by decreasing the production cost of the raw material, that is, lactic acid. It is of great importance to isolate and develop robust and highly efficient microbial lactic acid producers. Alongside the fermentative substrate and concentration, the yield and productivity of lactic acid are key parameters and major factors in determining the final production cost of lactic acid. In this review, we will discuss the current limitations and challenges for cost-efficient microbial production of optically pure lactic acid. The main obstacles to effective fermentation are the use of food resources, indirect utilization of polymeric sugars, sensitivity to inhibitory compounds released during biomass treatments, substrate inhibition, decreased lactic acid yield and productivity, inefficient utilization of mixed sugars, end product inhibition, increased use of neutralizing agents, contamination problems, and decreased optical purity of lactic acid. Furthermore, opportunities to address and overcome these limitations, either by fermentation technology or metabolic engineering approaches, will be introduced and discussed.

L-Lactic acid production from glycerol coupled with acetic acid metabolism by Enterococcus faecalis without carbon loss.

Glycerol is a by-product in the biodiesel production process and considered as one of the prospective carbon sources for microbial fermentation including lactic acid fermentation, which has received considerable interest due to its potential application. Enterococcus faecalis isolated in our laboratory produced optically pure L-lactic acid from glycerol in the presence of acetic acid. Gas chromatography-mass spectrometry analysis using [1, 2-(13)C2] acetic acid proved that the E. faecalis strain QU 11 was capable of converting acetic acid to ethanol during lactic acid fermentation of glycerol. This indicated that strain QU 11 restored the redox balance by oxidizing excess NADH though acetic acid metabolism, during ethanol production, which resulted in lactic acid production from glycerol. The effects of pH control and substrate concentration on lactic acid fermentation were also investigated. Glycerol and acetic acid concentrations of 30 g/L and 10 g/L, respectively, were expected to be appropriate for lactic acid fermentation of glycerol by strain QU 11 at a pH of 6.5. Furthermore, fed-batch fermentation with 30 g/L glycerol and 10 g/L acetic acid wholly exhibited the best performance including lactic acid production (55.3 g/L), lactic acid yield (0.991 mol-lactic acid/mol-glycerol), total yield [1.08 mol-(lactic acid and ethanol)]/mol-(glycerol and acetic acid)], and total carbon yield [1.06 C-mol-(lactic acid and ethanol)/C-mol-(glycerol and acetic acid)] of lactic acid and ethanol. In summary, the strain QU 11 successfully produced lactic acid from glycerol with acetic acid metabolism, and an efficient fermentation system was established without carbon loss.

Enterococcus faecium QU 50: a novel thermophilic lactic acid bacterium for high-yield l-lactic acid production from xylose.

Production of optically pure lactic acid from lignocellulosic material for commercial purposes is hampered by several difficulties, including heterofermentation of pentose sugars and high energy consumption by mesophilic lactic acid bacteria. Here, we report a novel lactic acid bacterium, strain QU 50, that has the potential to produce optically pure l-lactic acid (≥99.2%) in a homofermentative manner from xylose under thermophilic conditions. Strain QU 50 was isolated from Egyptian fertile soil and identified as Enterococcus faecium QU 50 by analyzing its sugar fermentation pattern and 16S rRNA gene sequence. Enterococcus faecium QU 50 fermented xylose efficiently to produce lactic acid over wide pH (6.0-10.0) and temperature ranges (30-52°C), with a pH of 6.5 and temperature of 50°C being optimal. To our knowledge, this is the first report of homofermentative lactic acid production from xylose by a thermophilic lactic acid bacterium.

Fed-batch fermentation for enhanced lactic acid production from glucose/xylose mixture without carbon catabolite repression.

There has been tremendous growth in the production of optically pure l-lactic acid from lignocellulose-derived sugars. In this study, Enterococcus mundtii QU 25 was used to ferment a glucose/xylose mixture to l-lactic acid. Maintenance of the xylose concentration at greater than 10 g/L achieved homo-lactic acid fermentation and reduced the formation of byproducts. Furthermore, carbon catabolite repression (CCR) was avoided by maintaining the glucose concentration below 25 g/L; therefore, initial concentrations of 25 g/L glucose and 50 g/L xylose were selected. Supplementation with 5 g/L yeast extract enhanced the maximum xylose consumption rate and consequently increased lactic acid production and productivity. Finally, a 129 g/L lactic acid without byproducts was obtained with a maximum lactic acid productivity of 5.60 g/(L·h) in fed-batch fermentation with feeding a glucose/xylose mixture using ammonium hydroxide as the neutralizing agent. These results indicate a potential for lactic acid production from glucose and xylose as the main components of lignocellulosic biomasses.

Recent advances in lactic acid production by microbial fermentation processes.

Fermentative production of optically pure lactic acid has roused interest among researchers in recent years due to its high potential for applications in a wide range of fields. More specifically, the sharp increase in manufacturing of biodegradable polylactic acid (PLA) materials, green alternatives to petroleum-derived plastics, has significantly increased the global interest in lactic acid production. However, higher production costs have hindered the large-scale application of PLA because of the high price of lactic acid. Therefore, reduction of lactic acid production cost through utilization of inexpensive substrates and improvement of lactic acid production and productivity has become an important goal. Various methods have been employed for enhanced lactic acid production, including several bioprocess techniques facilitated by wild-type and/or engineered microbes. In this review, we will discuss lactic acid producers with relation to their fermentation characteristics and metabolism. Inexpensive fermentative substrates, such as dairy products, food and agro-industrial wastes, glycerol, and algal biomass alternatives to costly pure sugars and food crops are introduced. The operational modes and fermentation methods that have been recently reported to improve lactic acid production in terms of concentrations, yields, and productivities are summarized and compared. High cell density fermentation through immobilization and cell-recycling techniques are also addressed. Finally, advances in recovery processes and concluding remarks on the future outlook of lactic acid production are presented.