25 USD. The manufacturing cost (consumables, including feedstock, labor, and utilities) would be approximately 2.35 USD/kg, and the total cost including taxes would be approximately 3.05 USD/kg. From the economic point of view, it is necessary to improve the production of propionic acid from apple pomace, to increase the yield of fermentation and thus decrease the total production costs. This can be achieved, for example, using industrial byproducts as nitrogen and vitamin sources, instead of high-cost substrates such as yeast extract or peptone. The online version contains supplementary material available at 10.1007/s13205-020-02582-x. The online version contains supplementary material available at 10.1007/s13205-020-02582-x.Sequential pretreatments for sugarcane bagasse (scb) by NaOH followed by organosolv under mild conditions were evaluated for cellulose recovery and dilignification. The best-optimized sequential pretreatment of scb was obtained at 10% (w/v) of raw scb loading at 1% (w/v) NaOH (50 °C, 2 h) followed by treatment with organosolv (85%, v/v phosphoric acid, 50 °C, 1 h) with chilled acetone. This sequentially pretreated scb showed cellulose recovery, 66.1% (w/w) and delignification, 83.2% (w/w). NaOH or organosolv pretreated scb showed lower cellulose recovery 47.4% (w/w) or 54.5% (w/w) with lower delignification, 61% (w/w) or 56% (w/w), respectively. Pretreated solid residue of sequentially pretreated scb was enzymatically saccharified by chimera (β-glucosidase and endoglucanase, CtGH1-L1-CtGH5-F194A) and cellobiohydrolase (CtCBH5A) cloned from Clostridium thermocellum. Enzymatic hydrolysate of best sequentially pretreated scb gave total reducing sugar (TRS) yield, 230 mg/g and glucose yield, 137 mg/g pretreated scb. Only organosolv pretreated scb gave TRS yield, 112.5 mg/g and glucose yield, 72 mg/g of pretreated scb. Thus, sequentially pretreated scb resulted in 37% higher enzymatic digestibility than only orgnaosolv pretreated scb. Higher enzymatic digestibility was supported by higher crystallinity index CrI (45%) than those obtained with only organosolv pretreated (38%) or raw scb (25%). Field Emission Scanning Electron Microscope (FESEM) and Fourier-transform infrared (FT-IR) analyses showed enhanced cellulose exposure in sequentially pretreated scb. Preliminary investigation of bioethanol production at small scale by separate hydrolysis and fermentation (SHF) of enzymatic hydrolysate from best sequentially pretreated scb by Saccharomyces cerevisiae gave maximum ethanol yield of 0.42 g/g of glucose. The online version contains supplementary material available at 10.1007/s13205-020-02600-y. The online version contains supplementary material available at 10.1007/s13205-020-02600-y.The present study was aimed to exploit the haloarchaeon Haloferax alexandrinus GUSF-1 (KF796625) for the presence of biomolecules possessing antioxidant activity. The culture produced a bright orange pigment when grown aerobically in nutrient rich medium with 25% crude solar salt. Biomolecules from cell-free supernatant and from the cells of the culture were individually extracted through the assistance of solvents of different polarities, such as ethanol, methanol and hexane, and monitored for scavenging of stable free radicals. Each of the extracts showed varying capacities to scavenge DPPH•(20, 31, and 80% DPPH• RSA; 160.19, 248.29 and 640.76 AAE µg g-1 of cells) at 1 mg mL-1. The extracellular ethanolic extract was polysaccharide in nature, equivalent to 47 µg mL-1 of glucose when assayed with the phenol-sulfuric acid method. The Fourier Transform-Infra Red spectroscopy confirmed the characteristic glycosidic peaks between 2000 and 1000 cm-1. Similarly, the glycerol diether moiety separated from hydroxylaerol dither moiety, tetrahydrosqualene, haloxanthin and 3-hydroxyechinenone is recorded as the first report for Haloferax alexandrinus GUSF-1 (KF796625). Therefore, recommended for use in microbial industrial biotechnology. The online version contains supplementary material available at 10.1007/s13205-020-02584-9. The online version contains supplementary material available at 10.1007/s13205-020-02584-9.In recent years, there has been an increasing interest in the remediation of contaminated environments, and a suitable solution is in situ bioremediation. To achieve this, large-scale bacterial biomass production should be sustainable, using economic culture media. The main aim of this study was to optimize the physicochemical conditions for the biomass production of an actinobacterium with well-known bioremediation ability using inexpensive substrates and to scale-up its production in a bioreactor. For this, the growth of four strains of actinobacteria were evaluated in minimal medium with glucose and glycerol as carbon and energy sources. In addition, l-asparagine and ammonium sulfate were assayed as alternative nitrogen sources. The strain Streptomyces sp. A5 showed the highest biomass production in shake-flasks culture using glycerol and ammonium sulfate as carbon and nitrogen sources, respectively. Factorial designs with five factors (glycerol concentration, inoculum size, pH, temperature, and agitation) were employed to optimize the biomass production of Streptomyces sp. A5. The maximum biomass production was obtained using 5 g L-1 of glycerol, 0.25 µL of inoculum, pH 7, 30 °C and 200 rpm. Finally, the production was successfully scaled to a 2 L stirred tank bioreactor. The online version contains supplementary material available at 10.1007/s13205-020-02588-5. The online version contains supplementary material available at 10.1007/s13205-020-02588-5.Despite its convenience and precision, CRISPR-based gene editing approaches still suffer from off-target effects and low efficiencies, which are partially rooted in Cas9, the nuclease component of the CRISPR/Cas9 system. In this study, we showed how mouse genome editing efficiency can be improved by constitutive and inheritable expression of Cas9 nuclease. For this goal, a transgenic mouse line expressing the Cas9 protein (Cas9-mouse) was generated. For in vitro assessment of gene editing efficiency, the Cas9-mice were crossed with the EGFP-mice to obtain mouse embryonic fibroblasts (MEF) expressing both EGFP and Cas9 (MEFCas9-EGFP). Transfection of these cells with in vitro transcribed (IVT) EGFP sgRNA or phU6-EGFPsgRNA plasmid led to robust decrease of Mean Fluorescent Intensity (MFI) to 8500 ± 1025 a.u. and 13,200 ± 1006 a.u. respectively. However, in the control group, in which the MEFEGFP cells were transfected with a pX330-EGFPsgRNA plasmid, the measured MFI was 16,800 ± 2254 a.u. https://www.selleckchem.com/ For in vivo assessment, the Cas9-zygotes at two pronuclei stage (2PN) were microinjected with a phU6-HhexsgRNA vector and the gene mutation efficiency was compared with the wild-type (WT) zygotes microinjected with a pX330-HhexsgRNA plasmid.