Effects of salinity, temperature and pH on the survival of the nemertean Procephalothrix simulus Iwata, 1952

The salinity, temperature and pH tolerance of Procephalothrix simulus Iwata, 1952, were experimentally studied. In hypo-media, the nemerteans could survive 96 h in 3.3‰ solution at 10 °C (median lethal salinity [LS₅₀] was not determined at this temperature), and 96 h LS₅₀ were 7.3‰ and 13.5‰ at 20 °C and 30 °C, respectively. In hyper-media, 96 h LS₅₀ values were 53.9‰, 47.1‰ and 41.4‰ at 10 °C, 20 °C and 30 °C, respectively. The trend of body weight changes in diluted media indicated that this nemertean is a volume regulator. During a 96-h exposure in media at 0 °C, worms were thanatoid but could recover if the temperature was gradually elevated to 20 °C. In thermal tolerance experiments, the nemertean survived 96 h in seawater of 30 °C, and worms suffered high mortalities when the temperature exceeded 32 °C. Present results suggest that the interaction of temperature and salinity on the lethal effects on P. simulus is significant (P < 0.05). Elevated temperature (range 10-30 °C) decreased the worm's solute tolerance, and elevated salinity (range 18-38‰) decreased the worm's thermal tolerance. The survival pH level for this nemertean ranged from 5.00 to 9.20.     

Cyclic CO(2) emissions during the high temperature pulse of fluctuating thermal regime in eye-pigmented pupae of Megachile rotundata

Megachile rotundata (Hymenoptera: Megachilidae), the primary pollinator used in alfalfa seed production, may need to be exposed to low-temperature storage to slow the insects' development to better match spring emergence with the alfalfa bloom. It has been demonstrated that using a fluctuating thermal regime (FTR) improves the tolerance of pupae to low temperatures. Carbon dioxide emission rates were compared between four different FTRs, all with a base temperature of 6 °C and a daily high-temperature pulse. Four different high-temperature pulses were examined, 15 or 25 °C for 2 h and 20 °C for 1 or 2 h. A subset of pupae at the FTR base temperature of 6 °C exhibited continuous gas exchange and, once ramped to 20 or 25 °C, shifted to cyclic gas exchange. As temperatures were ramped down from the high-temperature pulse to 6 °C, the pupae reverted to continuous gas exchange. The following conclusions about the effect of FTR on the CO₂ emissions of M. rotundata pupae exposed to low-temperature storage during the spring incubation were reached: 1) the high temperature component of the FTR was the best predictor of respiratory pattern; 2) neither pupal body mass nor days in FTR significantly affected which respiratory pattern was expressed during FTRs; 3) cyclic gas exchange was induced only in pupae exposed to temperatures greater than 15 °C during the FTR high temperature pulse; and 4) a two hour pulse at 25 °C doubled the number of CO₂ peaks observed during the FTR pulse as compared to a two hour pulse at 20 °C.     

Cold adaptation of the terrestrial isopod,Porcellio scaber,to subnivean environments

The terrestrial isopod, Porcellio scaber, was susceptible to subzero temperature: both freezing and chilling were injurious. The level of cold hardiness against chilling and freezing showed different patterns in their seasonal variation. The lower lethal temperature causing 50% mortality, an indicator of the tolerance to chilling, ranged from-1.37°C in August to-4.58°C in December. The whole body supercooling point, the absolute limit of freeze avoidance, was kept at about-7°C throughout the year. The winter decrease in lower lethal temperature was concomitant with an accumulation of low molecular weight carbohydrates which are possible protective reagents against chilling injury, whereas the less seasonally variable supercooling point seemed to be associated with the year-round presence of gut content. Food derivatives may act as efficient ice nucleators. The different trend in seasonal changes between lower lethal temperature and supercooling point may be related to the microclimate of the hibernacula in subnivean environments, where the winter temperature became lower than the lower lethal temperature in the summer active phase, but remained higher than the summer supercooling point.Abbreviations LLT₅₀ lower lethal temperature inducing 50% mortality - SCP supercooling point - T _a ambient air temperature - T _s soil surface temperature     

The importance of fluctuating thermal regimes for repairing chill injuries in the tropical beetle Alphitobius diaperinus (Coleoptera: Tenebrionidae) during exposure to low temperature

Abstract. In this study, the impact of acclimation (1 month at 15 °C vs. breeding at 30 °C) and fluctuating thermal regimes (daily transfers from low temperatures to various higher temperatures for 2 h) on the cold tolerance of the tropical beetle, Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) was examined. Acclimation increased significantly the duration of survival (Lt₅₀) at a constant 5 °C (7.7 ± 0.3 days to 9.7 ± 0.5 days). Survival of acclimated and nonacclimated beetles increased slightly at alternating temperatures of 5 °C/10 °C or 5 °C/15 °C. When daily transfer to 20 °C was applied, survival (Lt₅₀) was improved markedly (nonacclimated: 15.5 ± 0.7 days, acclimated: 19.6 ± 0.6 days). The higher temperatures may allow progressive repair of injuries, and the effects of chilling may be repaired completely at 25 and 30 °C, a phenomenon recorded here for the first time. It is estimated that the theoretical upper threshold of chill injury (Th) of nonacclimated beetles is 15.1 °C whereas it is shifted down to 11.2 °C in acclimated beetles, which might enable this temperature to allow effective repair of injury.     

Effects of fluctuating thermal regimes on cold survival and life history traits of the spotted wing Drosophila(Drosophila suzukii)

Drosophila suzukii is an invasive pest causing severe damages to a large panel of cultivated crops.To facili tate its biocontrol with stratcgies such as sterile or incompatible insect techniques,D.suzukid must be mass-produced and then stored and transported under low temperature.Prolonged cold exposure induces chill injuries that can be mitigated if the cold period is interrupted with short warming intervals,referred to as fluctuating thermal regimes(FTR).In this study,we tested how to optimally use FTR to extend the shelf life of D.suzukii under cold storage.Several FTR parameters were asessed:temperature(15,20,25℃),duration(0.5,1,2,3 h),and frequency(every 12,24,36,48 h)of warming intervals,in two wild-type lines and in two developmental stages(pupac and adults).Generally,FTR improved cold storage tolerance with respect to constant low temperatures(CLT).Cold mortality was lower when recovery temperature was 20℃ or higher,when duration was 2 h per day or longer,and when warming interruptions occurred frequently(every 12 or 24 h).Applying an optimized FTR protocol to adults greatly reduced cold mortality over long-term storage(up to 130 d).Consequences of FTR on fitness-related traits were also investigated.For adults,poststorage survival was unaffected by FTR,as was the case for female fecundity and male mating capacity.On the other hand,when cold storage occurred at pupal stage,postorage survival and male mating capacity were altered under CLT,but not under FTR.After storage of pupae,female fecundity was lower under FTR compared to CLT,suggesting an energy trade-off between repair of chill damages and C22 production.This study provides detailed information on the application and optimization of an FTR-based protocol for cold storage of D.suzuki that could be useful for the biocontrol of this pest.     

A comparison of low temperature tolerance traits between closely related aphids from the tropics, temperate zone, and Arctic

The survival of aphids exposed to low temperatures is strongly influenced by their ability to move within and between plants and to survive exposure to potentially lethal low temperatures. Little is known about the physiological and behavioural limitations on aphid movement at low temperatures or how they may relate to lethal temperature thresholds. These questions are addressed here through an analysis of the thermal ecology of three closely related aphid species: Myzus persicae, a ubiquitous temperate zone pest, Myzus polaris, an arctic species, and Myzus ornatus, a sub-tropical species. Lower lethal temperatures (LLT₅₀) of aphids reared at 15 °C were similar for M. persicae and M. polaris (range: −12.7 to −13.9 °C), but significantly higher for M. ornatus (−6.6 °C). The temperature thresholds for activity and chill coma increased with rearing temperature (10, 15, 20, and 25 °C) for all clones. For M. polaris and M. ornatus the slopes of these relationships were approximately parallel; by contrast, for M. persicae the difference in slopes meant that the difference between the temperatures at which aphids cease walking and enter coma increased by approximately 0.5 °C per 1 °C increase in rearing temperature. The data suggest that all three species have the potential to increase population sizes and expand their ranges if low temperature limitation is relaxed.     

Interspecific variation in the response to low temperature storage in different aphid parasitoids

Cold storage of natural enemies usually involves placing insects under constant subambient temperatures. Even at non-freezing temperatures, a reduction in survival is the norm. Using fluctuating thermal regimes (FTR) instead of constant low temperature (CLT) has shown that mortality due to accumulation of chilling injuries was significantly reduced in Aphidius colemani . Whether this phenomenon can be generalised to other parasitoid species is not known. The aim of this study was to analyse interspecific variation in the ability to tolerate cold storage under CLT (continuous 2°C) versus FTR (daily cycle: 2°C for 22 h and 20°C for 2 h) for various durations (0–20 days). Survival, sex ratio and development of five different Aphidiine parasitoids were analysed: A. colemani , Aphidius ervi , Aphidius matricariae , Ephedrus cerasicola and Praon volucre. A marked interspecific variation in the ability to tolerate cold storage was observed: A. matricariae and A. ervi were most chill tolerant, P. volucre and E. cerasicola had an intermediate chill sensitivity and A. colemani was most chill sensitive. In all species tested, FTR significantly reduced cold-induced mortality. This phenomenon was manifested more in chill-sensitive species as they probably accumulate chilling injuries more rapidly. The sex ratio remained unaffected in all the species. Interspecific variation was also observed in developmental responses to cold storage. Under CLT, time to adult emergence of A. matricariae, A. colemani, A. ervi and P. volucre was temporarily stopped and in E. cerasicola it increased. Under FTR, the short daily intervals at 20°C for 2 h allowed parasitoids to continue development in all the species. Interspecific differences are discussed. This study suggests that positive impact of FTR may apply to a wide range of species.     

Heat and starvation induced hormesis in longevity of Oomyzus sokolowskii (Kurdjumov) (Hymenoptera: Eulophidae) adult females

Oomyzus sokolowskii (Kurdjumov) (Hymenoptera: Eulophidae) is an important endoparasitoid of Plutella xylostella (Linnaeus) (Lepidoptera: Plutellidae) larvae and pupae. Previous studies have showed that environmental stress induced hormesis in a variety of organisms. In the present study, we investigated the effects of heat and starvation stress on the survival and induced hormesis in longevity of O. sokolowskii adult females under laboratory conditions. Results showed that temperature and exposure time significantly affected the survival rate of O. sokolowskii adult females with the extreme temperatures and/or longer durations proving to be more lethal. When O. sokolowskii adult females were heat-treated for 0.5, 1, 2, 4, and 8 h, LTemp₅₀ were >50.00, 43.24, 38.82, 38.32, and 38.17 °C, respectively. LTime₅₀ at the threshold temperature of 38 °C was 3.90 h. The sub-lethal temperature exposure for a certain time period reduced the survival numbers, but increased the longevity of survived adult females at the condition of starvation. The exposure for 2 h at 36 and 38 °C reduced the survival numbers by 26.67% and 46.67%, but extended the longevity of the survived adult females by 65.89% and 55.37% in comparison with those in the control, respectively. These results suggest O. sokolowskii adult female has the potential of thermal resistance, and its longevity will increase via heat and starvation treatment.     

Cold tolerance in rats following administration of streptozotocin,glucose, insulin and glucose plus insulin

Fall in rectal temperature (T_re) and survival time was determined on exposure to–20°C in adult normoglycemic and diabetic (streptozotocin treated) rats and 1 h following glucose feeding or insulin administration or both and on exposure to–10°C in young rats with and without glucose feeding. The susceptibility to frostbite was determined by exposure of the limbs to freezing mixture of–19°C or–23°C. The rate of fall of T_re was less and the survival time more in glucose and insulin plus glucose treated animals. On the other hand, the rate of fall of T_re was more and the survival time less, in dia betic and insulin-treated animals. The rectal temperature at which the animal died was the same in the control and the treated animals. The susceptibility to frost bite was more in insulin treated and diabetic animals and less in glucose-fed animals. Exposure to cold during the second h after glucose or glucose plus insulin injection did not alter the blood glucose from that obtained at room temperature. In insulin-treated animals the rate of rise of blood glucose during the second h was much higher at low temperature than at room temperature. The rise in blood glucose in diabetic animals was much higher than in normoglycemic animals exposed to cold.     

Effect of cultivation temperature,clomiphene, and nystatin on the oxidation and cyclization of chanoclavine in submerged cultures of the mutant strainclaviceps purpurea 59

Submerged cultures ofClaviceps purpurea 59 produce predominantly tricyclic chanoclavine and chanoclavine-I-aldehyde; formation of the tetracyclic agroclavine and elymoclavine is limited to 20%. The production cultures were characterized by the oxidation index o_i (100% chanoclavine/% chanoclavine) and cyclization index c_i (% agroclavine +% elymoclavine/% chanoclavine-I-aldehyde), expressing two functions of chanoclavine cyclase. Both indices were influenced by cultivation temperature and membrane agents. At 20°–24°C, clomiphene increased o_i and c_i; at 28°C and more it increased o_i only. At 24°C nystatin increased o_i and decreased c_i. During cultivation of vegetative inocula and production cultures at various temperatures (24°, 28°, and 33°C), the lower cultivation temperature of the inoculum increased c_i of the production culture with the higher temperature. The higher cultivation temperature of the inoculum decreased o_i and particularly c_i of the production culture with the lower temperature. In production cultures cultivated from the inoculum of an identical temperature, o_i decreased to 50% with increasing temperature above 24°C, whereas c_i decreased continuously. The role of the chanoclavine cyclase conformation as a membrane enzyme is discussed.     

Thermal sensitivity of heart rate and insensitivity of blood pressure in the Antarctic nototheniid fish <Emphasis Type="Italic">Pagothenia borchgrevinki</Emphasis>

The Antarctic notothenioids are among the most stenothermal of fishes, well adapted to their stable, cold and icy environment. The current study set out to investigate the thermal sensitivity/insensitivity of heart rate and ventral aortic blood pressure of the Antarctic nototheniid fish Pagothenia borchgrevinki over a range of temperatures. The heart rate increased rapidly from –1 to 6°C (Q₁₀=2.0–3.3), but was relatively insensitive to temperature above the ~6°C lethal limit of the species (Q₁₀=1.2). The increase in heart rate from –1 to 6°C was the result of a 45% increase in excitatory adrenergic tone, masking a 37% increase in inhibitory cholinergic tone. Ventral aortic pressure was regulated well above the lethal limit, up to at least 10°C. With the return of the fish to environmental temperatures, the heart rate rapidly decreased back to control levels, while ventral aortic pressure increased and remained elevated for over an hour following a 6°C exposure.     

Modelling time‐varying low‐temperature‐induced mortality rates for pupae of Tuta absoluta (Gelechiidae,Lepidoptera)

In a series of laboratory experiments, acclimated pupae of Tuta absoluta were exposed to various constant low temperatures in order to estimate their maximum survival times (Kaplan–Meier, Lt_99.99). A Weibull function was fitted to the data points, describing maximum survival time as a function of temperature. In another experiment at ?6°C, the progress of mortality increasing with exposure time was identified. These values were fitted by a sigmoidal function converging asymptotically to 100% mortality for very long exposure times. Analysing mortality data from the maximum survival experiment by a generalized linear model showed a significant common slope parameter (p < .001) that reveals parallelism of the survival curves at each temperature if a log time axis is used. These curves appear stretched (time scaled) if plotted with a nonlogarithmic time axis. By combining these mathematical relations, it was possible to calculate a species‐specific ‘mortality surface’ which exhibits mortalities, depending on temperature and duration of exposure. In order to accumulate hourly mortalities for courses of varying temperatures, an algorithm was developed which yields mortality values from that surface taking into account the attained mortality level. In validation experiments, recorded mortalities were compared against modelled mortalities. Prediction of mortality was partially supported by the model, but pupae experiencing intensely fluctuating temperatures showed decreased mortality, probably caused by rapid cold hardening during exposure. Despite this observation, mortality data converged to distinct levels very close to 100% depending on the intensity of temperature fluctuations that were characteristic for different types of experiments. The highest mortality limit occurred at intensely fluctuating temperatures in laboratory experiments. This constituted a benchmark that was not reached under various field conditions. Thus, it was possible to identify temperature limits for the extinction of field populations of Tuta absoluta pupae.     

Temperature and salinity tolerances of larval Californian grunion, Leuresthes tenuis (Ayres): A comparison with gulf grunion, L. Sardina (Jenkins & Evermann)

Temperature and salinity tolerances were determined for larval California grunion, Leuresthes tenuis (Ayres), and compared with previous data for Gulf of California grunion, L. sardina (Jenkins & Evermann). Larvae of similar age and acclimation history showed little interspecific difference in thermal tolerance, as measured by half-hour LT₅₀ values for 20–30 day old late postlarvae acclimated at various temperatures, and by upper and lower incipient lethal temperatures for 18°C-acclimated prolarvae. The upper incipient lethal temperature differed by 1 deg.-C (32°C for L. tenuis, 31°C for L. sardina), while the lower incipient lethal temperature for the 18°C acclimated prolarvae of both species was 7.5°C. L. tenuis larvae were much less euryhaline than L. sardina, with incipient lethal salinities of 4.2–41 %. for prolarvae and 8.6–38 %. for 20-day-old postlarvae; comparable values for L. sardina are 4–67.5 %. and 5–57.5 %. Both species show a decrease in temperature and salinity tolerance with age. The larvae of these disjunct congeners show a significant physiological divergence in euryhalinity but not in overall temperature tolerance. These tolerances are discussed in relation to the respective geographic ranges and behavioral responses of the two species.     

Induction of freezing tolerance in alfalfa cell suspension cultures

The effects of ABA, 2,4-D, kinetin and cold exposure on the cold hardiness of Medicago sativa L. cell suspensions were investigated. Cultures treated with 5×10^–5 M ABA at 2°C for 4 weeks in the absence of kinetin showed a 50% survival after freezing to –12.5°C, whereas cultures grown at 25°C under normal conditions tolerated freezing to only –3°C. The optimum ABA treatment of 5×10^–5 M for 4 weeks was effective only in combination with cold exposure. Of six cell lines tested, all showed different degrees of induced cold hardiness. The results suggest that ABA alone cannot induce freezing tolerance on alfalfa cell suspension cultures and that the deletion of kinetin and combination of low temperature and ABA is critical for the induction of cold hardiness in alfalfa cell suspension cultures.Abbreviations ABA abscisic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - LT₅₀ 50% killing temperature     

Rapid cold hardening increases cold and chilling tolerances more than acclimation in the adults of the sycamore lace bug, Corythucha ciliata (Say) (Hemiptera: Tingidae)

The sycamore lace bug, Corythucha ciliata is a new, invasive pest of Platanus trees in China. Although C. ciliata is often subjected to acute low temperatures in early winter and spring in northern and eastern China, the cold tolerance of C. ciliata has not been well studied. The objectives of this study were to determine whether adults of C. ciliata are capable of rapid cold hardening (RCH), and to compare the benefits of RCH vs. cold acclimation (ACC) in the laboratory. When the adult females incubated at 26 °C were transferred directly to the discriminating temperature (−12 °C) for 2 h, survival was only 22%. However, exposure to 0 °C for 4 h before transfer to −12 °C for 2 h induced RCH, i.e., increased survival to 68%. RCH could also be induced by gradual cooling of the insects at rates between 0.1 and 0.25 °C min⁻¹. The protection against cold shock obtained through RCH at 0 °C for 4 h was lost within 1 h if the adults were returned to 26 °C before exposure to −12 °C. Survival at both −12 and −5 °C was greater for RCH-treated than for ACC-treated adults (for ACC, adults were kept at 15 °C for 5 days), and the lethal temperature (2 h exposure) was lower for RCH-treated than for ACC-treated adults. The results suggest that RCH may help C. ciliata survive the acute low temperatures that often occur in early winter and early spring in northern and eastern China.     

Rapid thermal responses and thermal tolerance in adult codling moth Cydia pomonella (Lepidoptera: Tortricidae)

In order to preserve key activities or improve survival, insects facing variable and unfavourable thermal environments may employ physiological adjustments on a daily basis. Here, we investigate the survival of laboratory-reared adult Cydia pomonella at high or low temperatures and their responses to pre-treatments at sub-lethal temperatures over short time-scales. We also determined critical thermal limits (CTLs) of activity of C. pomonella and the effect of different rates of cooling or heating on CTLs to complement the survival assays. Temperature and duration of exposure significantly affected adult C. pomonella survival with more extreme temperatures and/or longer durations proving to be more lethal. Lethal temperatures, explored between −20 °C to −5 °C and 32 °C to 47 °C over 0.5, 1, 2, 3 and 4 h exposures, for 50% of the population of adult C. pomonella were −12 °C for 2 h and 44 °C for 2 h. Investigation of rapid thermal responses (i.e. hardening) found limited low temperature responses but more pronounced high temperature responses. For example, C. pomonella pre-treated for 2 h at 5 °C improved survival at −9 °C for 2 h from 50% to 90% (p < 0.001). At high temperatures, pre-treatment at 37 °C for 1 h markedly improved survival at 43 °C for 2 h from 20% to 90% (p < 0.0001). We also examined cross-tolerance of thermal stressors. Here, low temperature pre-treatments did not improve high temperature survival, while high temperature pre-treatment (37 °C for 1 h) significantly improved low temperature survival (−9 °C for 2 h). Inducible cross-tolerance implicates a heat shock protein response. Critical thermal minima (CTmin) were not significantly affected by cooling at rates of 0.06, 0.12 and 0.25 °C min⁻¹ (CTmin range: 0.3-1.3 °C). By contrast, critical thermal maxima (CTmax) were significantly affected by heating at these rates and ranged from 42.5 to 44.9 °C. In sum, these results suggest pronounced plasticity of acute high temperature tolerance in adult C. pomonella, but limited acute low temperature responses. We discuss these results in the context of local agroecosystem microclimate recordings. These responses are significant to pest control programmes presently underway and have implications for understanding the evolution of thermal tolerance in these and other insects.     

昆虫耐寒性的测定与评价方法

昆虫耐寒性因其重要的理论和实践意义成为当前生态学研究中一个热点领域。近年来 ,大量的文献从不同的角度报道了昆虫耐寒性及其生态学意义。该文根据近期昆虫耐寒性研究的文献并结合作者的实验结果 ,从研究方法的角度概述昆虫耐寒性实验研究中应注意的几个问题 ,包括持续低温和瞬间低温 ,恒定低温和变化的低温 ,以及冷却速率对昆虫耐寒性的影响。同时介绍评价昆虫耐寒性的几个参数和模型 ,并对耐寒性的定量表达和评价方法进行讨论。     

Physiological and biochemical analysis of overwintering and cold tolerance in two Central European populations of the spruce bark beetle, Ips typographus

Overwintering success is one of the key aspects affecting the development and outbreaks of the spruce bark beetle, Ips typographus (L.) populations. This paper brings detailed analysis of cold tolerance, and its influence on overwintering success, in two Central European populations of I. typographus during two cold seasons. Evidence for a supercooling strategy in overwintering adults is provided. The lower lethal temperature corresponds well to the supercooling point that ranges between −20 and −22 °C during winter months. The supercooled state is stabilized by the absence of internal ice nucleators and by seasonal accumulation of a mixture of sugars and polyols up to the sum concentration of 900 mM. The cryoprotective function of accumulated metabolites is probably based on increasing the osmolality and viscosity of supercooled body fluids and decreasing the relative proportion of water molecules available for lethal formation of ice nuclei. No activity of thermal hysteresis factors (stabilizers of supercooled state) was detected in hemolymph. Lethal times for 50% mortality (Lts50) in the supercooled state at −5, −10 or −15 °C are weeks (autumn, spring) or even months (winter), suggesting relatively little mortality caused by chill injury. Lts50 at −15 °C are significantly shorter in moist (6.9 days) than in dry (>42 days) microenvironment because there is higher probability of external ice nucleation and occurrence of lethal freezing in the moist situation.     

The recovery, penetrance, and pleiotropy of X-linked, cold sensitive mutants in Drosophila

Summary The viability at 16°, 22°, and 30°C, loci, visible phenotypes if any, possible effective lethal phase, and female fertility of seven X-linked, recessive cold sensitive mutations are reported. Five of the seven are female sterile at the restrictive temperature of 16°C; two of these five are also female sterile at the permissive temperature of 25°C. For two of the five mutations which are female sterile, escapers at the restrictive temperature exhibit visible phenotypes characteristic of mutants which affect protein synthesis. The possibility that some of the mutants affect ribosomes is considered. One of the mutants, l(1)TW-6 ^cs , is probably a cold sensitive meiotic mutant as well as a cold sensitive zygotic lethal. One of the mutants is a non-conditional visible allele of lozenge.Supported by NSF Grants GB 7707 and GB 20910.     

Biosynthesis of dextrans with different molecular weights by selecting the concentration of Leuconostoc mesenteroides B-512FMC dextransucrase, the sucrose concentration, and the temperature

Leuconostoc mesenteroides B-512FMC dextransucrase was found to synthesize dextrans of varying molecular weights by selecting the concentrations of dextransucrase and sucrose, as well as the temperature. Four enzyme concentrations (50, 10, 1.0, and 0.1 U/mL), five sucrose concentrations (20, 50, 100, 200 and 1000 mM), and two temperatures (20 °C and 30 °C) were studied. The highest amount of enzyme (50 U/mL), with the lowest concentration of sucrose (20 mM), and the lower temperature of 20 °C gave the lowest number-average molecular weight (MW_n) of 20,630 Da, respectively. As the sucrose concentration was increased, 50 mM, 100 mM, and 200 mM, the MW_n was 49,240 Da, 63,350 Da, and 126,720 Da, respectively. The next enzyme concentration (10 U/mL) gave a similar upward trend, starting at 73,130 Da and ending at 237,870 Da at 20 °C and 130,040 Da and ending at 415,770 Da at 30 °C. The upward trend continued for the 1.0 and 0.1 U/mL enzyme concentrations. An increase in the temperature had the overall effect of increasing the MW_n for each decreasing concentration of enzyme and increasing concentration of sucrose. For 0.1 U/mL and 1000 mM sucrose at 30 °C, the MW_n was 1,645,700 Da. The results of the study show that the molecular weights of the synthesized dextrans were inversely proportional to the concentration of the enzyme and directly proportional to the concentration of sucrose and the temperature.