The effect of light, salinity, and nitrogen availability on lipid production by Nannochloropsis sp

We examined responses of batch cultures of the marine microalga Nannochloropsis sp. to combined alterations in salinity (13, 27, and 40 g/l NaCl) and light intensity (170 and 700 μmol photons/m²·s). Major growth parameters and lipid productivity (based on total fatty acid determination) were determined in nitrogen-replete and nitrogen-depleted cultures of an initial biomass of 0.8 and 1.4 g/l, respectively. On the nitrogen-replete medium, increases in light intensity and salinity increased the cellular content of dry weight and lipids due to enhanced formation of triacylglycerols (TAG). Maximum average productivity of ca. 410 mg TFA/l/d were obtained at 700 μmol photons/m²·s and 40 g/l NaCl within 7 days. Under stressful conditions, content of the major LC-PUFA, eicosapentaenoic acid (EPA), was significantly reduced while TAG reached 25% of biomass. In contrast, lower salinity tended to improve major growth parameters, consistent with less variation in EPA contents. Combined higher salinity and light intensity was detrimental to lipid productivity under nitrogen starvation; biomass TFA content, and lipid productivity amounted for only 33% of DW and ca. 200 mg TFA/l/day, respectively. The highest biomass TFA content (ca. 47% DW) and average lipid productivity of ca. 360 mg TFA/l/day were achieved at 13 g/l NaCl and 700 μmol photons/m²·s. Our data further support selecting Nannochloropsis as promising microalgae for biodiesel production. Moreover, appropriate cultivation regimes may render Nannochloropsis microalgae to produce simultaneously major valuable components, EPA, and TAG, while sustaining relatively high biomass growth rates.     

Outdoor microalgae cultivation in airlift photobioreactor at high irradiance and temperature conditions: effect of batch and fed-batch strategies,photoinhibition, and temperature stress

The microalgae Scenedesmus abundans cultivated in five identical airlift photobioreactors (PBRs) in batch and fed-batch modes at the outdoor tropical condition. The microalgae strain S. abundans was found to tolerate high temperature (35–45 °C) and high light intensity (770–1690 µmol m^− 2 s^− 1). The highest biomass productivities were 152.5–162.5 mg L^− 1 day^− 1 for fed-batch strategy. The biomass productivity was drastically reduced due to photoinhibition effect at a culture temperature of > 45 °C. The lipid compositions showed fatty acids mainly in the form of saturated and monounsaturated fatty acids (> 80%) in all PBRs with Cetane number more than 51. The fed-batch strategies efficiently produced higher biomass and lipid productivities at harsh outdoor conditions. Furthermore, the microalgae also accumulated omega-3 fatty acid (C18:3) up to 14% (w/w) of total fatty acid at given outdoor condition.

     

Effects of Chromium and Carnitine Co-supplementation on Body Weight and Metabolic Profiles in Overweight and Obese Women with Polycystic Ovary Syndrome: a Randomized,Double-Blind,Placebo-Controlled Trial

The primary aim of our study was to determine the influence of taking chromium plus carnitine on insulin resistance, with a secondary objective of evaluating the influences on lipid profiles and weight loss in overweight subjects with polycystic ovary syndrome (PCOS). In a 12-week randomized, double-blind, placebo-controlled clinical trial, 54 overweight women were randomly assigned to receive either supplements (200 μg/day chromium picolinate plus 1000 mg/day carnitine) or placebo (27/each group). Chromium and carnitine co-supplementation decreased weight (− 3.6 ± 1.8 vs. − 1.0 ± 0.7 kg, P < 0.001), BMI (− 1.3 ± 0.7 vs. − 0.3 ± 0.3 kg/m², P < 0.001), fasting plasma glucose (FPG) (− 5.1 ± 6.0 vs. − 1.1 ± 4.9 mg/dL, P = 0.01), insulin (− 2.0 ± 1.4 vs. − 0.2 ± 1.2 μIU/mL, P < 0.001), insulin resistance (− 0.5 ± 0.4 vs. − 0.04 ± 0.3, P < 0.001), triglycerides (− 18.0 ± 25.2 vs. + 5.5 ± 14.4 mg/dL, P < 0.001), total (− 17.0 ± 20.3 vs. + 3.6 ± 12.0 mg/dL, P < 0.001), and LDL cholesterol (− 13.3 ± 19.2 vs. + 1.4 ± 13.3 mg/dL, P = 0.002), and elevated insulin sensitivity (+ 0.007 ± 0.005 vs. + 0.002 ± 0.005, P < 0.001). In addition, co-supplementation upregulated peroxisome proliferator-activated receptor gamma (P = 0.02) and low-density lipoprotein receptor expression (P = 0.02). Overall, chromium and carnitine co-supplementation for 12 weeks to overweight women with PCOS had beneficial effects on body weight, glycemic control, lipid profiles except HDL cholesterol levels, and gene expression of PPAR-γ and LDLR. Clinical trial registration number: http://www.irct.ir: IRCT20170513033941N38.

     

A submerged culture system for rapid micropropagation of the commercially important aquarium plant, ‘Amazon sword’ (Echinodorus ‘Indian Red’)

Echinodorus ‘Indian Red’ is an underwater plant, used worldwide for aquarium ornamentation. An efficient method for in vitro propagation and plantlet acclimatization of this popular aquarium plant was standardized. Surface-disinfected shoot-tips were cultured in submerged conditions in a solid–liquid bilayer medium, consisting of an upper, liquid layer (sterile distilled water) and a lower, solid layer Murashige and Skoog (MS) basal medium supplemented with 3.0% (w/v) sucrose, 0.8% (w/v) agar-agar, and plant growth regulators (PGRs) in different combinations and concentrations. The combination of 2.5 mg L⁻¹ 6-benzylaminopurine and 1.0 mg L⁻¹ α-naphthaleneacetic acid improved the multiplication rate to a maximum of 26.8 ± 0.51 shoots per explant after 60 d of culture. The number of multiplied shoots increased with each regeneration cycle, thus from only 26.8 ± 0.51 shoots per explant (first regeneration cycle), this number increased to 33.5 ± 0.58 (second regeneration cycle), and to 38.3 ± 0.62 for the third regeneration cycle with the same medium composition. The highest number of roots (8.3 ± 0.28) per shoot was induced in the presence of 1.0 mg L⁻¹ indole-3-butyric acid, but further growth of these roots was stunted. The best rooting was achieved on PGR-free ½-strength MS medium, where 6.1 ± 0.21 roots per shoot were induced with 5.8 ± 0.35 cm length after 30 d of culture. The regenerated plantlets were successfully acclimatized to submerged underwater conditions, with 100% survival rate. The present protocol is suitable for the commercial propagation of Echinodorus ‘Indian Red’ for aquarium-industries.

     

Saccharification of Parthenium hysterophorus biomass using cellulase from Streptomyces sp. NAA2

Parthenium hysterophorus biomass can be used as a non-conventional renewable feedstock for the production of bioethanol. Therefore, the present work was designed to hydrolyze P. hysterophorus biomass using cellulase enzyme produced from an actinomycete, i.e., Streptomyces sp. NAA2 using P. hysterophorus biomass as a substrate. The isolate NAA2 was identified by molecular characterization of 16SrDNA. The enzyme production by strain NAA2 was enhanced by optimization studies conducted under submerged fermentation conditions using P. hysterophorus as a substrate. The crude enzyme produced under optimized conditions was used to hydrolyze alkali-acid pretreated P. hysterophorus biomass. The highest CMCase production was achieved in 4–5 days when steam-pretreated P. hysterophorus biomass was used at 1% (w/v) concentration, using 2 discs (1 disc = 5 × 10⁷ spores/ml) of inoculum, an initial pH 6.5, temperature at 40 °C, an agitation speed of 120–150 rpm, and by supplementing fermentation medium with 1.5% (w/v) carboxymethyl cellulose (CMC) as additional carbon source. Under optimized conditions, the actinomycete strain NAA2 showed production of 0.967 ± 0.016 U/ml CMCase, 0.116 ± 0.08 FPU/ml FPase, and 0.22 ± 0.012 U/ml β-glucosidase enzymes. On utilizing the cellulase enzyme for biomass hydrolysis, maximum 18.2% saccharification yield (of cellulose 0.202 g/g) was achieved in 96 h when enzyme and substrate levels were 30 FPU/100 ml and 2% (w/v) respectively. Parthenium hysterophorus biomass can be hydrolyzed enzymatically yielding considerable amounts of total reducing sugars. It can, therefore, be used as a feedstock for the production of bioethanol. Also, it has the potential to act as a substrate for the production of cellulases. Furthermore, the improved cellulolytic potential of Streptomyces sp. NAA2 can be exploited in various industrial applications.

     

Microbial lipids from cellulolytic oleaginous fungus Penicillium citrinum PKB20 as a potential feedstock for biodiesel production

Microbial lipids derived from oleaginous fungi are considered as an alternative feedstock for biodiesel production. We attempt to isolate a cellulolytic oleaginous fungi as a potential feedstock for biodiesel production. The fungus was identified by 5.8 S-ITS rRNA gene sequencing. The extracellular enzyme activities were recorded after every 24 h for 7 days. Nile red staining and fluorescence microscopy was used to visualise the lipid bodies within the fungal hyphae. A renewable heterogeneous base catalyst derived from Musa balbisiana cola peels was used for the transesterification of Penicillium citrinum PKB20 derived oil into biodiesel. GC-MS analysis was used to analyse the fatty acid methyl esters (FAME) profile of the transesterified lipids. Penicillium citrinum PKB20 was isolated from detritus rich soil of Assam, India. The endoglucanase, xylanase and β-glucosidase enzyme activities were found to be 292.83 ± 0.29, 111.72 ± 0.45 and 6.54 ± 0.13 U/mg respectively. The specific enzyme activity for extracellular lipase was found to be 3.12 ± 0.16 U/mg. It could accumulate up to 60.61% of lipids in nitrogen-limited medium (7.34 ± 0.45 g/L biomass production). The extracted lipids were converted to biodiesel with 89.3% conversion efficiency. The predominant fatty acids were oleic acid (30.09%), palmitic acid (20.25%) and linoleic acid (33.14%) suggesting a balance between oxidative stability and cold flow properties for suitable biodiesel quality. Penicillium citrinum PKB20 was found to be a potential feedstock for biodiesel production with desirable fuel properties. The cellulolytic nature could be utilised for simultaneous lipid production directly on cellulosic substrates.     

Impact of In-Situ CO2 Nano-Bubbles Generation on Freezing Parameters of Selected Liquid Foods

The impact of in-situ CO₂ nano-bubbles generation on the freezing properties of soft serve, milk, and apple juice was investigated. Carbonated (0, 1000, and 2000 ppm) liquid foods contained in a tube were submerged and cooled for 90 min in a pre-set ethylene glycol bath (−15 °C). Before the enclosed liquid reached 0 °C, the vibration was discharged through ultrasound in the bath to create nano-bubbles within the carbonated food samples, and the changes in temperature for 90 min of each food were recorded as a freezing curve. The time for onset of nucleation of control soft serve mix was halved in samples with 2000-ppm CO₂ due to the presence of nano-bubbles. Likewise, the nucleation time for milk with and without nano-bubbles at the same CO₂ concentration of 2000 ppm was 7.9 ± 0.1 and 2.8 ± 0.8 min_, respectively. The generation of CO₂ nano-bubbles from 2000-ppm CO₂ level in 10 ^oBx apple juice displayed −9.3 ± 0.3 °C nucleation temperature while the control one had −11.7 ± 0.9 °C.

     

The Effects of Probiotic Honey Consumption on Metabolic Status in Patients with Diabetic Nephropathy: a Randomized,Double-Blind,Controlled Trial

To the best of our knowledge, this study is the first evaluating the effects of probiotic honey intake on glycemic control, lipid profiles, biomarkers of inflammation, and oxidative stress in patients with diabetic nephropathy (DN). This investigation was conducted to evaluate the effects of probiotic honey intake on metabolic status in patients with DN. This randomized, double-blind, controlled clinical trial was performed among 60 patients with DN. Patients were randomly allocated into two groups to receive either 25 g/day probiotic honey containing a viable and heat-resistant probiotic Bacillus coagulans T11 (IBRC-M10791) (10⁸ CFU/g) or 25 g/day control honey (n = 30 each group) for 12 weeks. Fasting blood samples were taken at baseline and 12 weeks after supplementation to quantify glycemic status, lipid concentrations, biomarkers of inflammation, and oxidative stress. After 12 weeks of intervention, patients who received probiotic honey compared with the control honey had significantly decreased serum insulin levels (− 1.2 ± 1.8 vs. − 0.1 ± 1.3 μIU/mL, P = 0.004) and homeostasis model of assessment-estimated insulin resistance (− 0.5 ± 0.6 vs. 0.003 ± 0.4, P = 0.002) and significantly improved quantitative insulin sensitivity check index (+ 0.005 ± 0.009 vs. − 0.0007 ± 0.005, P = 0.004). Additionally, compared with the control honey, probiotic honey intake has resulted in a significant reduction in total-/HDL-cholesterol (− 0.2 ± 0.5 vs. + 0.1 ± 0.1, P = 0.04). Probiotic honey intake significantly reduced serum high-sensitivity C-reactive protein (hs-CRP) (− 1.9 ± 2.4 vs. − 0.2 ± 2.7 mg/L, P = 0.01) and plasma malondialdehyde (MDA) levels (− 0.1 ± 0.6 vs. + 0.6 ± 1.0 μmol/L, P = 0.002) compared with the control honey. Probiotic honey intake had no significant effects on other metabolic profiles compared with the control honey. Overall, findings from the current study demonstrated that probiotic honey consumption for 12 weeks among DN patients had beneficial effects on insulin metabolism, total-/HDL-cholesterol, serum hs-CRP, and plasma MDA levels, but did not affect other metabolic profiles. http://www.irct.ir: IRCT201705035623N115.

     

Somatic embryogenesis and plant regeneration of <Emphasis Type="Italic">Lilium ledebourii</Emphasis> (Baker) Boiss., an endangered species

A somatic embryogenesis (SE) protocol was established for the regeneration of Lilium ledebourii (Baker) Boiss. whole plants using new vegetative bulblet microscales and transverse thin cell layers (tTCLs) of young bulblet roots as the explant sources. Bulblets were induced from bulb scale explants cultured for at least 3 months in the dark on Murashige and Skoog (MS) medium containing 3% sucrose, 0.8% agar, and different concentrations of α-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron. Embryo-like structures were obtained from tTCL explants of 3-month-old bulblets (excised from bulb scale explants) following culture on solid MS medium containing 3% sucrose and various concentrations of NAA and BA for 3 months in the dark. Both the explant source and the type of plant growth regulators affected the differentiation of somatic embryos. The highest percentage (65.55%) of embryogenesis was obtained from bulblet microscale tTCLs cultured on solid MS medium containing 0.54 μM NAA and 0.44 μM BA. Plants with normal shoots and roots were obtained following a 3-month culture of embryos on growth regulator-free MS medium at 25 ± 1°C under a 16/8-h light/dark photoperiod (light intensity 40 μmol m⁻² s⁻¹, cool-white fluorescent light). The plants were successfully acclimatized in the growth chamber.     

Designing next generation recombinant protein expression platforms by modulating the cellular stress response in Escherichia coli

Resistance towards known antimalarial drugs poses a significant problem, urging for novel drugs that target vital proteins in the malaria parasite Plasmodium falciparum. However, recombinant production of malaria proteins is notoriously difficult. To address this, we have investigated two putative K+ channels, PfKch1 and PfKch2, identified in the P. falciparum genome. We show that PfKch1 and PfKch2 and a C-terminally truncated version of PfKch1 (PfKch11−1094) could indeed be functionally expressed in vivo, since a K+-uptake deficient Saccharomyces cerevisiae strain was complemented by the P. falciparum cDNAs. PfKch11−1094-GFP and GFP-PfKch2 fusion proteins were overexpressed in yeast, purified and reconstituted in lipid bilayers to determine their electrophysiological activity. Single channel conductance amounted to 16 ± 1 pS for PfKch11−1094-GFP and 28 ± 2 pS for GFP-PfKch2. We predicted regulator of K+-conductance (RCK) domains in the C-terminals of both channels, and we accordingly measured channel activity in the presence of Ca2+.     

Attached culture of Nannochloropsis oculata for lipid production

The influences of urea, nitrate and glycine with four concentration levels on attached culture of Nannochloropsis oculata were investigated. The organic nitrogen source glycine was effective on improving not only adhesion biomass productivity but also adhesion rate. The maximum adhesion biomass productivity of 15.76 ± 0.52 g m^?2 day^?1 with adhesion rate of 76.67 ± 0.42 % was achieved with 18 mM glycine. To increase the lipid production, three lipid enhancing strategies were conducted afterwards, including nitrogen starvation, high light, and the combination of nitrogen starvation and high light. In nitrogen starvation situation, although the lipid content was greatly increased, the adhesion biomass productivity dropped probably due to the low cell viability. Increasing light intensity was effective on enhancing both adhesion biomass productivity and lipid content. The results indicated that nitrogen starvation was effective on improving both lipid content and adhesion rate when high light was applied. The maximum lipid yield of 4.32 ± 0.14 g m^?2 day^?1 with adhesion biomass productivity of 21.32 ± 0.65 g m^?2 day^?1, adhesion rate of 86.81 ± 0.10 % and lipid content of 20.24 ± 0.06 % was achieved with the combination strategy.     

Characterization of an extracellular polysaccharide produced by a Saharan bacterium Paenibacillus tarimensis REG 0201M

The purpose of this paper is to highlight the novel molecular characteristics and rheological properties of the polysaccharide produced by a bacterium isolated from extremophilic environment (Algerian Sahara). Phenotypic and molecular characteristics of the REG 0201M bacterial strain retained for this research were studied. Extracellular polysaccharide produced by REG 0201M was purified, characterized and its production was investigated. Analysis of 16S rDNA gene sequence showed that strain REG 0201M belonged to the species Paenibacillus tarimensis. In sucrose agar medium, 1.31 g dry weight of the polysaccharide per liter was produced. Evaluation of water absorption capacity showed that this polysaccharide was able to absorb 1000 times more water than its own weight. The average molecular weight determined by high-performance size exclusion chromatography multiangle laser light scattering was 1.718 × 106 g mol−1. Analysis of the monosaccharide composition by high-performance anion exchange chromatography showed the presence of fructose (77.67%) as the main neutral sugar, followed by galactose (20.37%), arabinose (1.79%), and rhamnose (0.16%). Its global charge determined by the Zeta potential measurement was about −35.27 ± 0.66 mV. The main functional groups were elucidated by Fourier-Transform Infrared Spectroscopy while the surface morphology was resolved by scanning electron microscopy analysis. Moreover, the rheological data revealed shear thinning properties with the same general behavior of the studied polysaccharide compared to xanthan. These results show the great potential of this polysaccharide, which could help to promote its use in various industries by replacing synthetic polymers.     

Effects of Initial Phosphorus Concentration and Light Intensity on Biomass Yield per Phosphorus and Lipid Accumulation of Scenedesmus sp. LX1

Phosphorus has been considered as one of the most important limiting resources of large-scale production of microalgal biofuel. The approaches to increase biomass yield per phosphorus, along with the lipid accumulation properties of Scenedesmus sp. LX1, were investigated in this study. It was found that practical biomass yield per phosphorous was reduced with the increase of initial phosphorus (P) concentration, but increased with light intensity. The highest biomass yield per P of 4,500 kg-biomass/kg-P was achieved at initial phosphorus concentration of 0.05 mg?·?L^?1 under the light intensity of 320 μmol photon?·?m^?2?·?s^?1. Furthermore, the lipid content per biomass and triacylglycerols (TAGs) content per lipid were found to be positively correlated to biomass yield per P. With the biomass yield increased from 2,800 kg-biomass/kg-P to 4,500 kg-biomass/kg-P, the lipid content per microalgal biomass and TAG content per lipid increased from 18.7 % to 35.0 % and from 69.5 % to 83.0 %. These results suggested a possible approach to achieve high biomass production and high lipid content simultaneously.     

Development of an efficient micropropagation system for Tecoma stans (L.) Juss. ex Kunth using thidiazuron and effects on phytochemical constitution

The present study was designed to investigate the stimulatory effects of different doses (0.1 to 2.5 μM) of thidiazuron (TDZ) on in vitro shoot induction and proliferation of mature nodal explants of Tecoma stans. Of the tested concentrations, 2.0 μM TDZ proved to be optimal for maximum regeneration (91%) with a mean shoot number of 5.6 ± 0.67, and length of 2.38 ± 0.08 cm, after 4 wk of incubation. To determine the negative effects of prolonged TDZ exposure, after 4 wk of incubation at optimized level of TDZ, the cultures were transferred to a secondary medium either lacking plant growth regulators or supplemented with benzyladenine (BA) alone, or in combination with different auxins (indole-3-acetic acid, indole-3-butyric acid, or α-naphthalene acetic acid; NAA). Among the tested concentrations, 2.5 μM BA in combination with 0.5 μM NAA yielded the maximum mean shoot number (16.60 ± 0.40), and average shoot length (4.76 ± 0.15 cm) after 4 wk of culture. The best rhizogenesis (93%) was achieved on ½ MS medium containing 1.5 μM NAA, with a mean root number of 7.60 ± 0.40 and length of 4.11 ± 0.23 cm, after 4 wk of incubation. The micropropagated plantlets were successfully acclimatized and hardened off in Soilrite™ with a 90% survival rate. The plantlets grew well with normal growth, flowering and showed, by gas chromatography–mass spectroscopy, an increase in the number of bioactive compounds compared with the donor plant. This is the first report on T. stans in vitro regeneration using TDZ.

     

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of <Emphasis Type="Italic">Bacopa monnieri</Emphasis> and accumulation of bacoside A in regenerated shoots

Brahmi (Bacopa monnieri) is an important medicinal plant mainly used for the treatment of neurological disorders and depression. Recent investigations revealed that bacoside A is major chemical component shown to be responsible for memory facilitating action of brahmi. The current investigation was carried out to assess the potential for increasing biomass and the concentration of bacoside A in the in vitro regenerated shoots by varying sucrose and pH levels of shoot regeneration medium. The leaf explants were cultured on the Murashige and Skoog (MS) medium supplemented with 2 mg l⁻¹ kinetin (KN) and with varying concentrations of sucrose (0, 1, 2, 3, 4, 5 and 6% at pH 5.8) and pH (4.5, 5.0, 5.5, 6.0 and 6.5 with 2% sucrose) with the objective of verifying the effects of sucrose and pH level on shoot regeneration and to verify the accumulation of bacoside A in the regenerated shoots. The shoot biomass increased (150.50 ± 2.84 shoots per explant, fresh wt 6.31 ± 0.12 g and dry wt 250 ± 5.00 mg) on the medium supplemented with 2% sucrose and pH which was set at 4.5. The results of HPLC analysis indicate that increase in sucrose concentration (0, 1, 2, 3, 4, 5 and 6% at pH 5.8) lead to decrease in the bacoside A content (39.51, 22.43, 13.05, 12.17, 10.73, 9.56 and 8.93 mg g⁻¹ dry wt, respectively) in regenerated shoots. These findings provide evidence that stressful condition of inadequate supply of carbon elevated synthesis of bacoside A in brahmi shoots. However, 2% sucrose is found suitable for biomass accumulation. Therefore, medium supplemented with 2% sucrose and pH set at 4.5 was found suitable for both biomass (6.31 ± 0.12 g fresh wt and 250 ± 5.00 mg dry wt) and bacoside A accumulation (13.09 mg g⁻¹ dry wt).     

Long-term (24h) cooling of ovarian fragments in the presence of permeable cryoprotectants prior to freezing: Two unsuccesful IVF-cycles and spontaneous pregnancy with baby born after re-transplantation

Cancer is the second major cause of death in the world. The problem of post-cancer infertility plays a significant role, because chemotherapy can be gonadotoxic. Cryopreservation of ovarian tissue before cancer therapy with re-implantation after convalescence is the potential key solution to this problem. The aim of this study was to test the viability of cryopreserved human ovarian cortex after long-term cooling in culture medium composed of permeable cryoprotectants. Ovarian fragments from sixteen patients were randomly divided into two groups. After the operation, tissue pieces assigned to both groups were cooled to 5 °C for 22–24 h, frozen and thawed. Group 1 pieces (n = 32) were cooled before cryopreservation in the standard culture medium, and Group 2 pieces (n = 32) were cooled in the freezing medium (culture medium+6% ethylene glycol+6% dimethyl sulfoxide+0.15 M sucrose). Freezing was performed in standard 5 ml cryo-vials with ice formation at −9 °C, cooling from −9 to −34 °C at a rate of −0.3 °C/min and plunging at −34 °C into liquid nitrogen. After thawing in a 100 °C (boiling) water bath, the removal of cryoprotectants was performed in 0.5 M sucrose with 20 min exposure in sucrose and 30 min stepping rehydration. The effectiveness of the pre-freezing cooling of tissue was evaluated by the development of follicles (histology). Six months after the autotransplantation, oocytes from the twenty-seven-year old, hormonally stimulated patient were retrieved and fertilized with her partner sperm through the intracytoplasmic spermatozoa injection (ICSI). For groups 1 and 2, 93.5 ± 1.9% and 96.4 ± 2.0% of the preantral follicles, respectively, were morphologically normal (P > 0.1) (with a tendency toward increasing in quality in Group 2). Six months after the auto-transplantation, two ICSI cycles resulted in the gathering and transplantation of high quality embryos, but no pregnancy had been established. Thirteen months after the auto-transplantation, the patient became spontaneously pregnant and delivered a healthy baby girl at term. Long-term (24 h) cooling of ovarian tissue to 5 °C before cryopreservation in the presence of permeable cryoprotectants simplifies the protocol of cryopreservation and has a tendency of increasing of the cells viability after thawing.     

Characterization,modification, and overexpression of 3-phosphoglycerate dehydrogenase in Corynebacterium glutamicum for enhancing l-serine production

The direct fermentative production of l-serine from renewable biomass using Corynebacterium glutamicum is attracting increasing attention. In this study, wild-type C. glutamicum SYPS-062 produced up to 6.65 ± 0.23 g/L l-serine; to further improve l-serine production, the serA gene was cloned, and the C-terminal domain of 3-phosphoglycerate dehydrogenase (PGDH) from this strain was truncated. When expressed in Escherichia coli, the resultant mutein SerAΔ197 showed a specific PGDH activity of 1.092 ± 0.05 U/mg protein, representing a decrease of 25.87 % from that encoded by serA, and was no longer sensitive to high concentrations of l-serine. When serA Δ591 was overexpressed in C. glutamicum SYPS-062, the activity of PGDH in C. glutamicum pJC1-tac-serA Δ591 increased by 47.72 %, and the resultant strain C. glutamicum pJC1-tac-serA Δ591 could accumulate 7.69 ± 0.22 g/L l-serine. Furthermore, when serA Δ591 was overexpressed in C. glutamicum SYPS-062ΔsdaA, the resultant strain could accumulate 8.84 ± 0.23 g/L l-serine at 102 h, and the yield of l-serine on cells (Y p/x) improved by 60 % when compared with that noted in the control. These results demonstrate that l-serine production in C. glutamicum SYPS-062 could be improved by overexpressing a C-terminal truncation of PGDH in combination with other genetic modifications.     

Production and partial characterization of the exopolysaccharide from Pleurotus sajor caju

Microbial exopolysaccharides (EPSs) are very important because they are used in biotechnological applications in different industrial areas. The aim of the study was to determine the best EPS producer Pleurotus sp., to optimize EPS production and to perform partial purification and characterization of the produced EPS. After the production conditions were optimized, the EPS was isolated and partially purified. EPS was characterized by HP-TLC, 1H-NMR, FT-IR, and TGA. Hydroxyl, superoxide, and DPPH radical scavenging activities of the EPS were also investigated spectrophotometrically. The best EPS producer and its incubation period in submerged fermentation were determined as Pleurotus sajor caju and on 5 days, respectively. Culture conditions to increase EPS production were optimized as follows (in per liter): 90 g of glucose, 10 g of yeast extract, 10 g of peptone, and 100 mM of Mg2+. The optimal initial pH, temperature, and an agitation rate of culture were determined as 5.0, 25 °C, and 150 rate min−1, respectively. The highest EPS production was determined as 33.32 ± 1.6 g L−1. After isolation of EPS, one active fraction was obtained by gel filtration chromatography. EPS is composed mainly of glucose according to HP-TLC analysis. To the results, EPS had a complex structure by having carbohydrate and protein contents. The produced EPS had high degradation temperature as well as high antioxidant activity.     

Growth and palmitoleic acid accumulation of filamentous oleaginous microalgae <Emphasis Type="Italic">Tribonema minus</Emphasis> at varying temperatures and light regimes

Palmitoleic acid (C16:1Δ9), contributes greatly to human health, industrial chemicals and biodiesel. The filamentous oleaginous microalgae Tribonema sp. has been identified as a highly efficient producer of palmitoleic acid. Temperature and light regime were adapted to regulate the palmitoleic acid content in this study. Strain T. minus was able to grow well at all the tested temperatures, even at 5 °C. The optimum temperature for palmitoleic acid accumulation (54.25 % of total fatty acid) was 25 °C. Moreover, both light intensity and photoperiod affect the growth, lipid content and fatty acid files of T. minus. The culture exposed to 240 μmol photons m^?2 s^?1 with a photoperiod of 24:0 showed the highest biomass (6.87 g L^?1) and biggest lipid content (61.27 % of dry weight), whereas the most amount of palmitoleic acid (50.47 % of total fatty acid) was detected at 120 μmol photons m^?2 s^?1. These findings make tangible contributions to culture T. minus for commercial production of lipid or palmitoleic acid.