Effect Of Cultivation Temperature Research Articles

Fungi of the Arctic Seas

The abundance and diversity of mycelial fungi in the bottom sediments of the Arctic Ocean seas (the Greenland, Barents and Kara seas) were studied. Samples of the surface bottom sediments were collected during the 84th (July‒August 2021) and 86th (October‒November 2021) cruises of RV Akademik Mstislav Keldysh. The taxonomic affiliation of the isolated fungi was determined using polyphasic taxonomy. The isolated fungi belonged to 16 genera of different classes of ascomycetous, basidiomycetous, and zygomycetous fungi. The effect of cultivation temperature and different NaCl concentrations on fungal growth was determined, as well as the effect of cultivation conditions on the fatty acid profile for the strains capable of growth on media with increased osmotic potential. While fatty acid composition was shown to be affected by changes in environmental conditions, the response to osmotic stress differed among the studied cultures from deep-sea sediments.

Simultaneous Production of MK-7 and Iturin A by Bacillus velezensis ND.

Bacillus velezensis can produce various secondary metabolites, such as the antibacterial compound iturin A and the coagulation-promoting menaquinone-7 (MK-7). To enhance the economic feasibility of the fermentation process, a co-production strategy, involving the simultaneous production of MK-7 and iturin A by Bacillus velezensis ND, was investigated in this study. Firstly, the effects of cultivation temperature and initial pH on the synthesis of MK-7 and iturin A were investigated. Considering the co-production of iturin A and MK-7, the optimal temperature and pH were determined as 32°C and 7, respectively. Subsequently, important nutrients for the co-production process were investigated. It was observed that glycerol, soybean meal, yeast extract, and L-glutamate had a significant effect on the co-produce process. An optimal medium composed of glycerol (72.19mL L-1), L-glutamate (1.4g L-1), yeast extract (16.88g L-1), and soybean meal (130.95g L-1) was obtained by response surface methodology (RSM). This co-produce process was further scaled up in a biofilm reactor, and the maximum concentration of MK-7 and iturin A reached 46.88mg L-1 and 5.58g L-1, respectively. Finally, we established an effective method for separately extracting the two metabolites from the fermentation broth. The superiority of this co-production fermentation strategy demonstrates its significant potential for industrial production.

Growth Temperature Influences Postharvest Quality and Cold Tolerance of Green Harvested Dwarf Tomatoes During Storage

Effect of cultivation temperature during the phase of flowering and fruit development on tomato quality was investigated. Plants of two dwarf tomato cultivars “Ponchi Re” and “Tarzan,” were subjected to three different growth temperatures: 16, 22, or 28°C, starting at the flowering phase. Mature green fruit was harvested and subjected to shelf life at 20°C for 20 days or first stored at 4°C for 15 days, and then placed under shelf life conditions. Fruit quality was determined through red color development, soluble solid content (SSC), softening, weight loss, and cold tolerance. Higher cultivation temperature increased development and production of fruit. Deviation from the 22°C growth temperature led to increased soluble solid content in both cultivars, and smaller fruit diameter in “Tarzan.” Fruit grown at lower temperature had delayed color development during shelf life, and this was further delayed by prior cold storage. “Tarzan” showed more chilling injury (CI) symptoms than “Ponchi Re.” In our experiment, SSC can be manipulated by modulating cultivation temperature, but that it is not associated with CI tolerance. Delayed color formation at the lowest growth temperature observed in “Ponchi Re” tomatoes could be resulted in lower lycopene levels leads to lower ROS scavenging capacity. For “Tarzan” tomatoes, higher firmness at harvest, less softening, and lower weight loss during cold storage in fruit from the lowest cultivation temperature might positively correlated with increased membrane integrity, resulting in increased CI tolerance. This indicates that CI incidence depends on growth temperature and is cultivar dependent in dwarf tomato fruit.

Co-Expression of Chaperones for Improvement of Soluble Expression and Purification of An Anti-HER2 scFv in Escherichia Coli.

Single-chain fragment variable (scFv) is one of the most commonly used antibody fragments. They offer some advantages over full-length antibodies, including better penetration to target tissues. However, their functional production has been a challenge for manufacturers due to the potential misfolding and formation of inclusion bodies. Here we evaluated the soluble expression and purification of molecular chaperone co-expression. E. coli BL21(DE3) cells were co-transformed with the mixture of plasmids pKJE7 and pET22b-scFv by the electroporation method. First, L-arabinose was added to induce the expression of molecular chaperones, and then IPTG was used as an inducer to start the expression of anti-HER2 scFv. The effect of cultivation temperature and IPTG concentration on soluble expression of the protein with or without chaperones was evaluated. The soluble expressed protein was subjected to native purification using the Ni-NTA affinity column. SDS-PAGE analysis confirmed the successful co-expression of anti-HER2-scFv and DnaK/DnaJ/GrpE chaperones. Co-expression with chaperones and low-temperature cultivation synergistically improved the soluble expression of anti-HER2 scFv. Co-expression with chaperone also exhibited an approximately four-fold increase in the final yield of purified soluble protein. The combination of co-expression with chaperones and low temperature presented in this work may be useful for the improvement of commercial production of other scFvs in E. coli as functionally bioactive and soluble form.

The Effects of Foliar Application of Phenoxy and Imidazoline Family Herbicides on the Limitation of Primary Photosynthetic Processes in Galega orientalis L.

Fodder galega (Galega orientalis) is a perennial, wintering plant with great potential for agricultural development. The species has a large yield potential and exceptional adaptability to various environmental conditions. The sensitivity of G. orientalis to herbicides, however, as well as the photosynthetic performance of the species, are generally unknown. Our study aimed to evaluate the effects of the application of selected phenoxy herbicides (MCPA, MCPB) and the imidazoline family herbicide (IMA) on the parameters of primary photosynthetic processes as understood through fast chlorophyll fluorescence kinetics (OJIP). The effect of cultivation temperature was also investigated in the plants grown at 5, 18 and 25 °C. Time courses of OJIP-derived parameters describing photosystem II functioning after foliar application revealed that the plants showed negative responses to the herbicides in the order MCPB–MCPA–IMA within 24 h after the application. The application of herbicides decreased the values of maximum chlorophyll fluorescence (FM) and increased minimum fluorescence (F0), which led to a reduction in the maximal efficiency of PSII (FV/FM). Applications of MCPA and MCPB decreased variable chlorophyll fluorescence at 2 ms (VJ), 30 ms (VI) and VP, as well as the performance index (PIABS), which is considered a vitality proxy. The application increased absorption flux (ABS/RC), trapped energy flux (TRo/RC) and dissipated energy flux (DIo/RC). The effects were more pronounced in plants grown at 18 and 25 °C. The study revealed that the OJIP-derived parameters sensitively reflected an early response of G. orientalis to the foliar application of herbicides. Negative responses of PSII were more apparent in MCPA- and MCPB- exposed plants than IMA-exposed plants.

Influence of Cultivation Conditions on the Sioxanthin Content and Antioxidative Protection Effect of a Crude Extract from the Vegetative Mycelium of Salinispora tropica.

Due to their bioavailability, glycosylated carotenoids may have interesting biological effects. Sioxanthin, as a representative of this type of carotenoid, has been identified in marine actinomycetes of the genus Salinispora. This study evaluates, for the first time, the effect of cultivation temperature (T) and light intensity (LI) on the total cellular carotenoid content (TC), antioxidant activity (AA) and sioxanthin content (SX) of a crude extract (CE) from Salinispora tropica biomass in its vegetative state. Treatment-related differences in TC and SX values were statistically significantly and positively affected by T and LI, while AA was most significantly affected by T. In the S. tropica CE, TC correlated well (R2 = 0.823) with SX and somewhat less with AA (R2 = 0.777). A correlation between AA and SX was found to be less significant (R2 = 0.731). The most significant protective effect against oxidative stress was identified in the CE extracted from S. tropica biomass grown at the highest T and LI (CE-C), as was demonstrated using LNCaP and KYSE-30 human cell lines. The CE showed no cytotoxicity against LNCaP and KYSE-30 cell lines.

The effect of cultivation temperature on the soybean (Glycine max) growth and community structure of bradyrhizobia

The effect of cultivation temperature on the soybean (<i>Glycine max</i>) growth and community structure of bradyrhizobia

Impact of maturation and growth temperature on cell-size distribution, heat-resistance, compatible solute composition and transcription profiles of Penicillium roqueforti conidia

Penicillium roqueforti is a major cause of fungal food spoilage. Its conidia are the main dispersal structures of this fungus and therefore the main cause of food contamination. These stress resistant asexual spores can be killed by preservation methods such as heat treatment. Here, the effects of cultivation time and temperature on thermal resistance of P. roqueforti conidia were studied. To this end, cultures were grown for 3, 5, 7 and 10 days at 25 °C or for 7 days at 15, 25 and 30 °C. Conidia of 3- and 10-day-old cultures that had been grown at 25 °C had D56-values of 1.99 ± 0.15 min and 5.31 ± 1.04 min, respectively. The effect of cultivation temperature was most pronounced between P. roqueforti conidia cultured for 7 days at 15 °C and 30 °C, where D56-values of 1.12 ± 0.05 min and 4.19 ± 0.11 min were found, respectively. Notably, D56-values were not higher when increasing both cultivation time and temperature by growing for 10 days at 30 °C. A correlation was found between heat resistance of conidia and levels of trehalose and arabitol, while this was not found for glycerol, mannitol and erythritol. RNA-sequencing showed that the expression profiles of conidia of 3- to 10-day-old cultures that had been grown at 25 °C were distinct from conidia that had been formed at 15 °C and 30 °C for 7 days. Only 33 genes were upregulated at both prolonged incubation time and increased growth temperature. Their encoded proteins as well as trehalose and arabitol may form the core of heat resistance of P. roqueforti conidia.

Optimization of glucoamylase production by Mucor indicus, Mucor hiemalis, and Rhizopus oryzae through solid state fermentation / Mucor indicus, Mucor hiemalis, ve Rhizopus oryzae tarafından üretien glukoamilazın katı hal fermantasyonu ile optimizasyonu

Abstract Objective: Glucoamylase is a hydrolyzing enzyme with several industrial applications. Glucoamylase was produced via a solid state fermentation by three naturally occurring zygomycetes fungi of Mucor indicus, Mucor hiemalis, and Rhizopus oryzae on wheat bran. Methods: The effects of cultivation temperature, medium moisture content, and cultivation time on the enzyme production were investigated. Experiments were designed with an orthogonal central composite design on the three variables using response surface methodology (RSM). Results: For glucoamylase production, the optimum temperature and medium moisture content for the three fungi were 26.6°C and 71.8%, respectively. The optimum cultivation time for M. hiemalis and R. oryzae was 33.1 h, while it was 66.8 h for M. indicus. At optimum conditions, glucoamylase production by M. indicus, M. hiemalis, and R. oryzae was respectively 255.3, 272.3, and 1545.3 U per g dry substrate. Conclusion: R. oryzae is a suitable candidate for industrial production of glucoamylase.

Effect of Environmental Conditions on the Expression Levels of a Recombinant 11S Amaranth Globulin in Escherichia coli

The expression in Escherichia coli strain Rosetta of the recombinant acidic subunit from the 11S amaranth seed storage protein (protein ACM3) was studied at flask and at bioreactor levels. This subunit was modified by inserting four Val-Tyr antihypertensive peptides in tandem into its third variable region and also with the tripeptide Ile-Pro-Pro in the Cterminal region. Flasks experiments allowed us to define the best conditions for the preparation and expression and accumulation of the protein ACM3, including the certainty of its presence within the cells especially as an insoluble fraction. The effects of cultivation temperature, aeration rate and agitation speed on the production of the protein ACM3 was tested in a 5-L batch bioreactor. Applying response surface methodology (RSM) we found that the aeration rate was the most significant factor affecting in a positively way the production yields and productivity of the recombinant protein. Temperature had effect only in conjunction to aeration. The highest recombinant acidic subunit concentration (747 mg L-1) and the highest productivity (186 mg L-1 h-1) were attained in 4 h of cultivation when the factors evaluated were controlled at its central values: 0.1 vvm, 300 rpm, and 30.5° C. Results from this study indicate that RSM is an effective technique to maximize the production of this recombinant protein.

メタン生成嫌気ベンゼン分解集積培養系に及ぼす有機酸と温度の影響

メタン生成嫌気ベンゼン分解集積培養系に及ぼす有機酸と温度の影響

Growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. under different cultivation temperature

Microalgal lipid is a promising feedstock for biodiesel production. Effect of cultivation temperature on the growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. LX1 was studied. Scenedesmus sp. LX1 could grow in a wide range of temperature (10∼30°C), and the growth activation energy was 49.3 kJ·mol(-1). The optimal temperature to produce microalgal biomass and lipid was 20°C, and after 15 days of batch cultivation the productivities of 313.3 g biomass·(g P)(-1), 112 g lipid (g P)(-1) and 14.7 g TAGs·(g P)(-1) were obtained. The content of polyunsaturated fatty acids decreased with the increase of cultivation temperature. The reactive oxygen species (ROS) levels at 10°C and 20°C were higher than that under higher temperature. For the first time the cultivation temperature, ROS level, specific growth rate and lipid content per microalgal biomass were correlated together.

Anaerobic Microbial Associations Degrading Aminoaromatic Acids

Anaerobic microbial associations have been isolated that degrade aminoaromatic acids to methane and carbon dioxide at high rates. Significant differences between the morphological, cytological, and physiological traits of cultures isolated from samples of adapted and unadapted sludge are shown. The effects of cultivation temperature, illumination, and presence of mineral nitrogen and bicarbonate in the medium upon adaptation of enrichment cultures to substrates and subsequent behavior of the anaerobic associations have been studied. Intermediate and final products of degradation of aminoaromatic compounds and the sequence of their formation in the cultures have been determined. We have also studied the effects of exogenous electron acceptors and additional carbon sources on the degradation of aminoaromatic compounds.

Quantitative Analysis of Ceramide III of Saccharomyces cerevisiae by Normal Phase HPLC

Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by normal phased high performance liquid chromatography (NP‐HPLC) using evaporative light scattering detector (ELSD) and mass spectrometer. The mobile phase was composed of chloroform and methanol. One linear gradient mode was applied to gain proper analysis of cell extracts. Quantitative analysis of ceramide was performed. Based on the analytical condition, the effect of cultivation temperature for the production of ceramide was investigated and the optimum cultivation temperature was found to be 30°C at pH 7. From the results of mass spectrometer analysis, C26 ceramide was the dominant cell culture. The fragment ions from the mass spectrum of ceramides III, which were of various molecular weights, from m/z = 237–677, were observed.

Optimization of agitation, aeration, and temperature conditions for maximum β-mannanase production

The effects of cultivation temperature, aeration rate, and agitation speed on the production of β-mannanase by Bacillus licheniformis NK-27 in a batch fermenter were investigated in this study. Results revealed that temperature was the most significant factor in terms of its effect on β-mannanase production. It influenced β-mannanase production by affecting the other parameters including bacteria growth, pH, dissolved oxygen, total and reducing sugars. Agitation speed and aeration rate could affect dissolved oxygen concentration which in turn affected cell growth and β-mannanase production. A maximum β-mannanase activity of 212.0 U ml −1 was attained in 36 h of cultivation when aeration rate, agitation speed, and temperature were controlled at 0.75 vvm, 600 rpm, and 30 °C, respectively. The maximum β-mannanase activity in the fermenter was close to that obtained from the shake flask fermentation study (198.2 U ml −1). However, the duration of fermentation cycle in the fermenter study was shorter than the corresponding duration obtained from the shake flask experiment by 12 h.

Production of cholesterol oxidase by Rhodococcus equi No. 23 in a jar fermenter.

Rhodococcus equi No. 23 was grown in a batch fermenter. The effects of cultivation temperature, pH of the culture medium, aeration rate and agitation speed on the production of cholesterol oxidase (CholOx) by the test organism were examined. Results revealed that the cultivation temperature, the pH of the medium, the aeration rate and the agitation speed all affected the production of CholOx by R. equi No. 23. Adjusting the operation variables during the cultivation period increased the production of CholOx effectively and prevented the occurrence of overflow of foam during the fermentation period. A maximum CholOx activity of 0.34 unit/ml with a volumetric production rate of 0.011 unit/h per ml could be achieved in 30 h of cultivation at an aeration rate of 5.0 l/min, if the pH of the culture medium, the cultivation temperature and the agitation speed were controlled at 6.5, 39 degrees C and 200 rev. /min respectively during the first 24 h of cultivation, then shifted to 7.5, 37 degrees C and 300 rev./min respectively.

バイオリアクターによる食酢製造工程の改善に関する研究 第５報 濃縮菌体の一部排出培養法による高酸度食酢の連続生産における培養温度の影響

To raise the acetic acid concentration and enhance productivity, the effect of cultivation temperature in the second fermentor was studied on continuous production of vinegar using hollow fiber module and two fermentors. ln the second fermentor, the cells were concentrated with a hollow fiber module and some portion of concentrated celis were discharged to suppress excessive increase in ce11 concentration. Cultivation temperature was regulated at 30°C in the first fermentor and varied from 20 to 30°C in the second fermentor. Fresh medium of an ethanol concentration of 13% (v/v) at a rate of 100 ml/h was fed. A broth containing 117 g-acetic acid/l of acetic acid and 0.114 g-dry cell/l of cell was produced in the first fermentor and supplied to the second fermentor. ln the second fermentor, cells were concentrated with a hollow fiber module and was discharged at a discharge ratio of 0.1. The acetic acid concentration reached 156 g-acetic acid/l at 24°C and was constant below 24°C. Total acetic acid production rate and the yield of both fermentors was 3.0 g-acetic acid (l·h) and 0.94 g-acetic acid/1.304 g-consumed ethanol respectively. In the acetic acid production, the ratio of contribution of the first and second fermentor was estimated as 0.72:0.28.

A new strategy for the cloning, overexpression and one step purification of three DHAP-dependent aldolases: Rhamnulose-1-phosphate aldolase, fuculose-1-phosphate aldolase and tagatose-1,6-diphosphate aldolase 1

Three DHAP-dependent aldolases, rhamnulose-1-phosphate aldolase (Rham-IPA), fuculose-1-phosphate aldolase (Fuc-1PA) and tagatose-1,6-diphosphate aldolase (TDPA) have been cloned and overexpressed in Escherichia coli using two different expression vectors: pTrcHis for the expression of Rham-1PA and Fuc-1PA and pRSET for the expression of TDPA. In each case the recombinant enzyme is synthesized as a fusion protein with a hexahistidine tag on the N-terminus. The three enzymes have been purified in only one step by chelation affinity chromatography. The effects of cultivation temperature and concentration of inducer have been studied in order to optimize the expression of the recombinant proteins and to avoid the formation of inclusion bodies.

Effect of Cultivation Temperature on the Spontaneous Development of Deletions in Soilborne Wheat Mosaic Furovirus RNA 2

Intraplant movement of soilborne wheat mosaic virus (SBWMV) and the spontaneous development of sequence deletions in SBWMV RNA 2 were monitored in individual plants of Triticum aestivum cultivars Galahad and Vona inoculated mechanically or with viruliferous zoospores of Polymyxa graminis and grown at different temperatures. Movement of SBWMV RNAs 1 and 2 (either full-length or deleted forms) from foliage to roots was detected 2 days after mechanical inoculation. Plants infected by P. graminis and maintained at high temperatures (25-30 C) showed extensive deletions in the readthrough (RT) domain of the coat protein-RT gene within 4- 12 wk. In contrast, plants kept at 17 C over the same period contained only full-length RNA 2 molecules (3,593 nucleotides). Thus, in addition to prolonged cultivation at 15- 17 C and / or serial mechanical transfers to healthy plants, higher temperatures resulted in truncated forms of SBWMV RNA 2. Our observations confirm that only full-length SBWMV RNA 2 is transmitted to wheat roots by viruliferous P. graminis from field soil and that there is no intraplant barrier to the movement of deleted forms of RNA 2 between roots and leaves. Deleted forms of SBWMV RNA 2 appear to cause more severe symptoms only after mechanical inoculation to young, healthy plants. The significance of these observations for events in fields of winter wheat or barley, especially those expressing a temperature-sensitive host resistance gene against SBWMV, are discussed. These studies may also help elucidate the mechanism of spontaneous RNA deletion

Effect of cultivation temperatures on thermophilic and mesophilic enzyme gene expression in Bacillus subtilis

Expression of the thermostable neutral protease gene, nprM, from Bacillus stearothermophilus MK232 was investigated at various temperatures, using Bacillus subtilis as a host strain. When the nprM gene was expressed at temperatures of 39°C or lower, enzyme production was observed in the declining phase of batch culture. In contrast, when the cultivation temperature was higher than 40°C, enzyme production started in the exponential growth phase and continued in to the declining phase. The enzyme production was enhanced with an increase in cultivation temperature. Maximum enzyme activity per unit cell mass reached 3,500 u/OD 600 at 50°C, which is twelve-fold higher than that at 30°C. When another thermostable neutral protease gene, nprT, from B. stearothermophilus CU21, was expressed in B. subtilis, enzyme production started in the exponential growth phase at 37 and 45°C. However, enzyme production decreased at higher temperatures. When B. subtilis a-amylase gene, amyE, was expressed in the same host cells, the enzyme production at 45°C was lower than that at 37°C. The different effects of cultivation temperature on the expression of some genes are discussed.