Overexpression of Shinorhizobium Meliloti Flavohemoglobinefficiently Improved Cell Growth and Fatty Acid Biosynthesis in Oleaginous Fungus Mucor Circinelloides

Oxygen availability is a limiting factor for lipid biosynthesis in eukaryotic microorganisms. Two bacterial hemoglobins from Vitreoscilla sp. (VHb) and Shinorhizobium meliloti (SHb), which could deliver the oxygen to the respiratory chain to produce more ATP, were introduced into Mucor circinelloides to alleviate oxygen limitation, thereby improving cell growth and fatty acid production. VHb and SHb genes were integrated into the M. circinelloides MU402 genome through homologous recombination, and their protein expression was verified by carbon monoxide difference spectrum (CO-difference spectrum)analysis. SHb-expressing strain showed higher biomass than VHb-expressing strain. The biomass of the SHb-expressing strain was increased by about 50% and the total fatty acid (TFA) content was as high as 15.7% of the dry cell weight which was about 40% higher than that of the control strain in flask conditions. In the fermenter, the maximum biomass and TFA content was obtained in SHb-expressing strains, with the biomass being 12.1 g/L and the TFA being 21.1% of the dry cell weight. VHb and SHb expression also affected the fatty acid composition with the proportion of polyunsaturated fatty acids being increased. Over-expression of bacterial hemoglobins, especially SHb increased cell growth and TFA content in M. circinelloides at low and high aeration, suggesting that SHb is better than VHb in improving the fatty acid production in oleaginous microorganisms.

Callus inducement of Toona sinensis: Potential agents against SARS-Corona virus replication

Bioprospecting of anti-SARS-corona virus phytochemical agents is an important issue today as an alternative to chemical drugs whose effectiveness has not been fully successful with no light effect. Quercetin, a component of Toona sinensis young leaves extract, was reported to have anti-viral activity against SARS-corona virus replication in vitro. Currently, the low efficiency of secondary metabolite production is an obstacle. Production of secondary metabolites from callus induction in vitro is considered more efficient and in a short time for commercial applications. The composition of hormones in callus media affects the secondary metabolites formed. The study was conducted to determine the effect of synthetic hormones auxin (BAP) and cytokinin (NAA) on callus percentage, fresh and dry callus cell weight and the average time of callus formation. The call percentage (100%) was obtained in different combinations of BAP and NAA in MS medium. The highest fresh weight of callus (165.50 gL−1) and dry cell weight (28.47 gL−1) were observed in MS medium (1.5 mgL−1 BAP and 1 mgL−1 NAA) within 7 days initial formation time of callus. The results showed that all the indices measured were positively correlated with callus induction in T. sinensis.

Effect of Feeding With Vitamins, Sugars and Pollen on The Biology of Honey Bee Apis Mellifera L. (Hymenoptera : Apidae) During The Autumn Season and The Use of Various Wintering Methods

The research included a study of nutrition with vitamins, sugars, pollen and some dispersal methods in from 5/7/2020 to 5/15/2021 to evaluate the nutrients and their effect on the open and closed brood space, honey area, pollen, eggs, the increase in cell weight, the quantity of honey produced as well as the number of packages of honey bee (Apis mellifera L) colonies. The results showed that feeding with vitamins, sugars and pollen has an important role in preparing the colonies before winter, as they passed it with strong colonies compare with the comparison treatment (without feeding) and with the highest numerical density in the nylon packing boxes treatment compared to the comparison treatment during the tenth month readings. The treatment of cork boxes in the fed colonies was distinguished in the production of immature honey compared to the control treatment. The results also showed the superiority of the colonies fed during the months of November and December 2020 with the same materials over the comparison treatment in bee density, open honey, sealed honey, pollen, eggs, larvae and sealed brood) compared to the control treatment. The fed and treated colonies passed some winterization requirements in good condition and gave the highest cell weights compared to the control treatment in mud boxes, sandwich boxes, wall coverings, cork boxes, and nylon packing. As for the production of sorted honey, the fed and dispersed colonies outperformed the fed and scattered colonies with treatments compared to the comparison treatment, and in the production of parcels, the comparison treatment did not produce any parcels, while the fed and dispersed colonies produced parcels.

Challenges and Potential in Increasing Lutein Content in Microalgae

Research on enhancing lutein content in microalgae has made significant progress in recent years. However, strategies are needed to address the possible limitations of microalgae as practical lutein producers. The capacity of lutein sequestration may determine the upper limit of cellular lutein content. The preliminary estimation presented in this work suggests that the lutein sequestration capacity of the light-harvesting complex (LHC) of microalgae is most likely below 2% on the basis of dry cell weight (DCW). Due to its nature as a structural pigment, higher lutein content might interfere with the LHC in fulfilling photosynthetic functions. Storing lutein in a lipophilic environment is a mechanism for achieving high lutein content but several critical barriers must be overcome such as lutein degradation and access to lipid droplet to be stored through esterification. Understanding the mechanisms underlying lipid droplet biogenesis in chloroplasts, as well as carotenoid trafficking through chloroplast membranes and carotenoid esterification, may provide insight for new approaches to achieve high lutein contents in algae. In the meantime, building the machinery for esterification and sequestration of lutein and other hydroxyl-carotenoids in model microorganisms, such as yeast, with synthetic biology technology provides a promising option.

Smart Production of Lipids as Bio-Fuel in Spirulina Platensis (=Arthrospira Fusiformis), and Bio-Oxygen and Bio-Electricity in Media Cultured in Supernatant of Digested Poultry Waste

An experiment was conducted to evaluate growth performances and production of bio-fuel of Spirulina platensis (Gomont), and bio-oxygen and bio-electricity of culture media in supernatant of three different amount of digested poultry waste (DPW), and Kosaric medium (KM) as control. Three different amounts (concentrations) such as 2.0, 4.0 and 6.0 g/L poultry waste were allowed to digest under aeration. After 17 days, 700 ml grayish coloured supernatant was taken with addition of 9.0 g/L NaHCO3 and 0.50 ml/l micronutrient in 2.0 L conical flask with three replications and then autoclaved. Spirulina was inoculated to grow in these three treatments including KM (Control) after 72 hours of autoclave and then allowed to grow for a period of 14 days. This duration was estimated through repeated growth trials. The cell weight of spirulina was attained a maximum of 12.58 ± 1.25 mg/L (dry wt. basis) in KM followed by 11.46 ± 1.03, 9.16 ± 0.84 and 8.13 ± 0.73 mg/L in supernatant of 4.0, 2.0 and 6.0 g/L DPW, respectively on the 10th day of culture. Similar trend was also observed in the cases of optical density of the media contained spirulina, chlorophyll a content (mg/L), cell weight, total biomass (mg/l) and specific growth rates on the basis of cell weight and chlorophyll a. Cell weight of spirulina grown in these media had highly significant (P < 0.01) correlation with the chlorophyll a content (r = 0.993) of spirulina. Crude protein of spirulina grown in supernatant of DPW was lower than that of spirulina cultured in KM. Crude lipids as bio-fuel of spirulina cultured in supernatant of 4.0 g/L DPW was almost three times higher than that of spirulina grown in KM which may be due to high phospholipids bioaccumulation. Phosphate-P was decreased in media of DPW due to its use for high lipids biosynthesis as phospholipids. Bio-oxygen was produced higher ranged from 2.1 ± 11 mg/L on initial day to 10.20 ± 0.54 mg/L on 10th day grown in supernatant of 4.0 g/L DPW media. Bio-electricity as green electricity ranged from 135 ± 4 on initial day to 284 ± 7 mV on 10th day when spirulina cultured in supernatant of 4.0 g/L DPW which was higher than grown in other media. pH followed the similar trend like bio-oxygen and bio-electricity. It was found that the production of bio-oxygen, bio-fuel and bio-electricity had direct relation with pH in culture media of spirulina. Therefore, mass production of Spirulina platensis might be done in supernatant of 4.0 g/L digested poultry waste to get high total lipids as bio-fuel, bio-electricity and bio-oxygen.

Smart Culture of Spirulina Using Supernatant of Digested Rotten Tomato (Solanum Lycopersicum) to Produce Protein, Bio-Fuel and Bio-Electricity

An experiment was conducted for the production of protein, bio-fuel and bio-electricity from the culture system of Spirulina platensis (Gomont) in supernatant of three different amount of digested rotten tomato (Solanum lycopersicum), and Kosaric Medium (KM) as control. Three different concentrations such as 25, 50 and 75% rotten tomato were allowed to digest under aeration. After 17 days, the colorless supernatant was screened and taken in 1.0 L conical flask with three replications. Then, Spirulina platensis was inoculated to grow in these three media (treatments) with the addition of 9.0 g/L NaHCO3 and micronutrients, and also in KM as control for a period of 14 days. The cell weight, optical density, chlorophyll a and total biomass of spirulina was attained to the maximum values when grew in KM on the 10th day of culture followed by supernatant of 50% digested rotten tomato (DRT) than in 25 and 75% DRT culture. The chemical properties of the culture media such as pH, salinity, dissolved bio-oxygen, electric conductivity and bio-electricity were increased from first day up to 12th day of experiment. Total biomass of spirulina grown in these media had highly significant (P < 0.01) correlation with cell weight (r = 0.825) and chlorophyll a (r = 0.866) of spirulina. The results showed that the growth performances of S. platensis grown in supernatant of 50% DRT was significantly (P < 0.01) higher than that of spirulina grown in supernatant of 25 and 75% DRT. The percentage of crude protein (55.10 ± 0.45 to 59.90 ± 0.33%) of spirulina grown in supernatant of DRT was little bit higher than that of spirulina cultured in KM (58.40 ± 0.38%). But bio-fuel in terms of crude lipids (16.50 ± 0.31%) of spirulina cultured in supernatant of 50% DRT was almost two and half times higher than that of spirulina grown in KM (crude lipids, 6.30 ± 0.21%). Bio-electricity (300 ±10.20 mV) produced in culture of spirulina in supernatant of 50% DRT was higher than that recorded in KM (240 ±10.20 mV) followed by 75% DRT and other media. Bio-electricity had directly and strongly significant (p < 0.001) correlation with pH (r = 0.812), dissolved bio-oxygen (r = 0.832), salinity (r = 0.788) and electric conductivity (r = 0.856). Therefore, this procedure will produce huge amount of electricity in the world and will make a revolution in this field of bio-electricity production. Whole world will be benefited from the output (results) of this experiment.

The Effects of Total Dissolved Carbon Dioxide on the Growth Rate, Biochemical Composition, and Biomass Productivity of Nonaxenic Microalgal Polyculture

The biosequestration of CO2 using microalgae has emerged as a promising means of recycling CO2 into biomass via photosynthesis, which could be used to produce biofuels as an attractive approach to CO2 mitigation. We investigated the CO2 fixation capability of the native nonaxenic microalgal culture using a 2 L photobioreactor operated in batch mode. The cultivation was carried out at varying concentrations of total dissolved CO2 (Tco2) in the bulk media ranging from 200 to 1000 mg L−1, and the temperature and light intensities were kept constant. A maximum CO2 fixation rate was observed at 400 mg L−1 of Tco2. Characteristic growth parameters such as biomass productivity, specific growth rate, maximum biomass yield, and biochemical parameters such as carbohydrate, protein, and lipids were determined and discussed. We observed that the effect of CO2 concentration on growth and biochemical composition was quite significant. The maximum biomass productivity was 22.10 ± 0.70 mg L−1 day−1, and the rate of CO2 fixation was 28.85 ± 3.00 mg L−1 day−1 at 400 mg L−1 of Tco2. The maximum carbohydrate (8.17 ± 0.49% dry cell weight) and protein (30.41 ± 0.65%) contents were observed at 400 mg L−1, whereas the lipid content (56.00 ± 0.82% dry cell weight) was the maximum at 800 mg L−1 of Tco2 in the bulk medium.

Perancangan Sistem Perangkap Hama Tanaman Petani Otomatis Menggunakan Modul Mikrokontroler Arduino

An animal can also be called a pest if it causes damage to natural ecosystems or acts as an agent for spreading disease in human habitats. The pest in question is a mouse-like animal that aims to make a trap using a load cell weight sensor with an Arduino microcontroller module. The scope of the problem in this research is the cultivation of maize and paddy fields in Mardingding District, Karo Regency. For testing purposes, 3 trials are used, respectively, corn and rice fields. The variable is limited to animals the size of a mouse. The purpose of this research is to build a plant pest trapping device using the Arduino uno micro controller module which can make it easier to catch pests efficiently. Farmers no longer need to control within a certain period of time. so it can save time and energy to do other jobs. The application is built using the programming language used is the C language and the Arduino IDE with notification to farmers only via short messages via cellphones.

Application of In-Situ and Soft-Sensors for Estimation of Recombinant P. pastoris GS115 Biomass Concentration: A Case Analysis of HBcAg (Mut+) and HBsAg (MutS) Production Processes under Varying Conditions

Microbial biomass concentration is a key bioprocess parameter, estimated using various labor, operator and process cross-sensitive techniques, analyzed in a broad context and therefore the subject of correct interpretation. In this paper, the authors present the results of P. pastoris cell density estimation based on off-line (optical density, wet/dry cell weight concentration), in-situ (turbidity, permittivity), and soft-sensor (off-gas O2/CO2, alkali consumption) techniques. Cultivations were performed in a 5 L oxygen-enriched stirred tank bioreactor. The experimental plan determined varying aeration rates/levels, glycerol or methanol substrates, residual methanol levels, and temperature. In total, results from 13 up to 150 g (dry cell weight)/L cultivation runs were analyzed. Linear and exponential correlation models were identified for the turbidity sensor signal and dry cell weight concentration (DCW). Evaluated linear correlation between permittivity and DCW in the glycerol consumption phase (<60 g/L) and medium (for Mut+ strain) to significant (for MutS strain) linearity decline for methanol consumption phase. DCW and permittivity-based biomass estimates used for soft-sensor parameters identification. Dataset consisting from 4 Mut+ strain cultivation experiments used for estimation quality (expressed in NRMSE) comparison for turbidity-based (8%), permittivity-based (11%), O2 uptake-based (10%), CO2 production-based (13%), and alkali consumption-based (8%) biomass estimates. Additionally, the authors present a novel solution (algorithm) for uncommon in-situ turbidity and permittivity sensor signal shift (caused by the intensive stirrer rate change and antifoam agent addition) on-line identification and minimization. The sensor signal filtering method leads to about 5-fold and 2-fold minimized biomass estimate drifts for turbidity- and permittivity-based biomass estimates, respectively.