Resistance and bioaccumulation of Cd2+, Cu2+, Co2+ and Mn2+ by thermophilic bacteria, Geobacillus thermantarcticus and Anoxybacillus amylolyticus

In this study, bioaccumulation and heavy metal resistance of Cd²⁺, Cu²⁺, Co²⁺ and Mn²⁺ ions by thermophilic Geobacillus thermantarcticus and Anoxybacillus amylolyticus was investigated. The bacteria, in an order with respect to metal resistance from the most resistant to the most sensitive, was found to be Mn²⁺ > Co²⁺ > Cu²⁺ > Cd²⁺ for both G. thermantarcticus and A. amylolyticus. It was determined that the highest metal bioaccumulation was performed by A. amylolyticus in Mn²⁺ (28,566 μg/g dry weight), and the lowest metal bioaccumulation was performed by A. amylolyticus in Co²⁺ (327.3 μg/g dry weight). The highest Cd²⁺ capacities of dried cell membrane was found to be 36.07 and 39.55 mg/g membrane for G. thermantarticus and A. amylolyticus, respectively, and the highest Cd²⁺ capacities of wet cell membrane was found to be 14.36 and 12.39 mg/g membrane for G. thermantarcticus and A. amylolyticus, respectively.     

Some implications of a constant fiber stress hypothesis in the diastolic left ventricle

A thick-wall incompressible, elastic sphere was used as a model for the diastolic rat left ventricle. A model for myocardial nonhomogeneity was derived assuming that fiber (circumferential) stress was independent of position in the ventricular wall. The theoretical implications of the resulting constitutive relations together with the spherical model were analyzed in the context of large deformation elasticity theory. It was found that muscle stiffness at a given level of uniaxial stress increased monotonically from the endocardium to the epicardium. In addition, fiber stress was found to be essentially a linear function of transmural pressure above a pressure of 6 g/cm². It was also shown theoretically that neglecting the nonhomogeneity of the myocardium resulted in a state of stress which differed significantly from that predicted by the nonhomogeneous model. For example, at a transmural pressure of 14 g/cm², fiber stress in the nonhomogenous model was equal to 17 g/cm² while fiber stress in the homogeneous model varied between 100 g/cm² at the endocardial surface and 2 g/cm² at the epicardial surface. The change in muscle stiffness with position which characterized the nonhomogeneous model also tended to linearize the highly curvilinear radial stress distribution predicted by the homogeneous model at a given transmural pressure.     

Optimized production of L-β-hydroxybutyric acid by a mutant of Yarrowia lipolytica

A mutant strain of Yarrowia lipolytica was developed which produced 8.0 g l--hydroxybutyric acid l^–1 from butyric acid in a batch culture. The optimum culture conditions in the fermenter for maintenance of a high cell activity, determined by chemostat analyses, were a specific growth rate of 0.06 h^–1, a glucose concentration of 2.0 g l^–1, and a butyric acid concentration of 8.1 g l^–1. A fed-batch fermentation was performed under these conditions resulting in an l--hydroxybutyric acid yield of 31 g l^–1.     

Isolation and characterization of insecticidal activity of (Z)-asarone from Acorus calamus L.

The insecticidal activity of Acorus calamus L. rhizome‐derived material against adults of Sitophilus zeamais Motschulsky was examined by using repellency method and contact toxicity. The biologically active constituent of the A. calamus rhizome was separated and identified. The results showed that the ethanol extract of A. calamus had strong repellency and contact effect to S. zeamais and the active constituent of the A. calamus was characterized as (Z)‐asarone by spectroscopic analysis. Responses from the tests varied with exposure times and doses. In the repellency test, ethanol extract of A. calamus had 93.92% repellency at 629.08 μg/cm² but only 71.38% at 157.27 μg/cm² 12 h after treatment. As a contrast, (Z)‐asarone showed 84.50% repellency at 314.54 μg/cm² and 77.02% at 78.63 μg/cm² 12 h after treatment. In the filter paper diffusion test, ethanol extract of A. calamus caused 95.56% and 17.78% mortality to S. zeamais at 314.54 μg/cm² and 78.63 μg/ cm² 4 days after treatment, while (Z)‐asarone brought about 100.00% and 15.56% mortality at 40.89 μg/cm² and 15.73 μg/cm² respectively. These results indicate that the insecticidal activity of the A. calamus extract may be due to (Z)‐asarone.     

Influence of the thermochemical sludge pretreatment on the nitrification of A/O reactor with the removal of phosphorus by simultaneous precipitation

In the present study, a laboratory scale anoxic/oxic (A/O) reactor is used for the removal of nutrient and sludge reduction. Phosphorus removal was achieved through simultaneous precipitation, and sludge production was reduced through thermochemical pretreatment. The main objective of the study was to investigate the influence of sludge pretreatment on the nitrification rate. Total phosphorus in the effluent was maintained around 0.5 ～ 1.0 mg/L by simultaneous precipitation, using coagulant alum at 2.2 mole ratio. Before simultaneous precipitation, the nitrification rate of the A/O reactor was found to be 0.050 g N-NH₄ ⁺/g MLVSS.d. The thermochemical sludge pretreatment began on the 120^th day at pH 11 and 80°C. The initiation of sludge pretreatment brought about a significant reduction of the A/O reactor nitrification rate, which fell to 0.038 g N-NH₄ ⁺/g MLVSS/day. The effect of sludge pretreatment was reflected in the reduction of the nitrogen removal efficiency from 85 to 74%. Recycling of the thermochemically pretreated sludge accounted for 57% sludge reduction, which had an adverse influence on the nitrification rate of the system.     

Continuous culture of Candida boidinii variant 60 growing on methanol

Summary A new variant, Candida boidinii variant 60, which is less sensitive to methanol and formaldehyde shocks was grown in continuous cultures with methanol as sole carbon source. The substrate concentration in the feeding medium was either 1% methanol or 3% methanol. Biomass production, methanol consumption, the formation of formaldehyde and gas exchange were measured at different dilution rates. With low methanol feeding (10 g/l) maximal productivity of 0.44 g biomass/l·h is obtained at a dilution rate of 0.14 h^–1. Maximal specific growth rate is 0.18 h^–1. A yield of 0.32 g biomass/g methanol was obtained and the respiration quotient was determined as 0.55. Independently of initial substrate concentration, biomass decreases if methanol and formaldehyde are accumulating in the culture broth.In the culture with high methanol feeding (30 g/l) cell concentratioon increases up to 9 g/l at D=0.04 h^–1. At higher dilution rates methanol and form-aldehyde appear in the medium. Formaldehyde is then preferably oxidized without energy advantages for the cells. It seems that this enables the cells to overcome toxic effects caused by methanol and formaldehyde.     

Continuous fermentation to produce fungal protein. Effect of growth rate on the biomass yield and chemical composition of Fusarium moniliforme

Fusarium moniliforme was grown on a carob aqueous extract in a chemostat for fungal protein production. The substrate was adjusted to provide 0.5% carob sugars supplemented with inorganic salts. The dilution rate varied from 0.086 to 0.227 hr^?1 under constant conditions of temperature (30°C), pH (4.5), and oxygen saturation (60–80%). A yield of 0.709 g dry mycelium/g consumed carob sugar and a productivity value of 0.687 g dry mycelium/liter hr^?1 were obtained at μ = 0.205 hr^?1. The maintenance coefficient was 0.077 g carob sugar/g dry mycelium hr^?1. While the carbohydrate and purine content of dry mycelium increased at μ values from 0.114 to 0.205 hr^?1 both true (Lowry) and crude (N × 6.25) protein contents decreased at the same μ range. Maximum values of 36.3% true and 47.9% crude protein of dry mycelium were obtained at μ = 0.114 hr^?1, whereas a minimum purine content of 99.8 μmol/g corresponding to 6.42% nucleic acids was recorded at μ = 0.086 hr^?1. It was concluded that a continuous fermentation of carob aqueous extract using F. moniliforme should be operated at growth rates of approximately 0.205 hr^?1 in order to maximize protein production.     

Removal of Styrene from Waste Gas Using a Biological Trickling Filter

Biodegradation of styrene in a biological trickling filter on lava stones was investigated, firstly, with the addition of silicon oil and, secondly, without the addition of silicon oil. After 400 days of trial runs the experimental results revealed that the biodegradation capacity of styrene in the trickling filter reached 537 g/m³ × h with a degradation yield of 96.8 % at an air inlet concentration of 1.06 g/m³ of styrene and a space velocity of 157 m/h in the presence of silicon oil. A removal of styrene up to 2.9 kg/m³ × h was obtained when the styrene input concentration in a constant inlet air flow of 0.78 m³/h was increased up to 6.6 g/m³. Interestingly, it was observed that after a period of 400 days, the seven dominant strains were completely different from those present in the inoculum. Surprisingly, this population was able to grow in an aqueous liquid phase without silicon oil on a styrene concentration of 45.5 g/L. In the biological trickling filter with lava stones but without silicon oil, the biodegradation capacity of styrene was 464 g/m³ × h with a removal yield of 98.3 % at an air inlet concentration of 1.03 g/m³ of styrene and a space velocity of 137 m/h. As in the presence of silicon oil, a removal of styrene of up to 2.375 kg/m³ × h was achieved when the air flow rate was kept constant and the styrene input concentration was increased. These experiments suggested that the biphasic medium could be very efficiently used for the selection of adapted strains for the removal of insoluble or poorly soluble organic compounds, rather than being used for long‐term degradation under industrial conditions.     

Peat accretion and phosphorus accumulation along a eutrophication gradient in the northern Everglades

Recent rates of peat accretion (as determined by¹³⁷Cs) and N, P, organic C, Ca and Na accumulation were measured along a 10 km eutrophication gradient in the northern Everglades area of Water Conservation Area 2A (WCA 2A) that has received agricultural drainage from the Hillsboro canal for the past 25–30 yrs. Rates of peat accretion were highest at sampling locations closest to the Hillsboro canal, 1.6 km downstream, (5.67 ± 0.50 mm/yr) and decreased to 2.01 ± 0.31 mm/yr at distances of 7.1 to 10.7 km downstream. Phosphorus and Na accumulation were a function of both peat accretion and soil P and Na concentrations. The concentration and accumulation of P in peat deposited in the past 26 years was highest near the Hillsboro canal (1478 ± 67 ug/g, 0.66 ± 0.06 g/m²/yr) and decreased to 560 ± 20 ug/g and 0.10 ± 0.02 g/m²/yr at distances of 8.8 to 10.7 km downstream. Like phosphorus, the concentration and rate of Na accumulation was highest near the Hillsboro canal (3205 ± 1021 ug/g, 1.48 ± 0.53 g/m²/yr). Although sodium enrichment of the peat was limited to 1.6 km downstream of the Hillsboro canal, increased rates of Na accumulation penetrated 5.2 km downstream of the Hillsboro canal, the extent of the area of enhanced peat accretion. In contrast to P and Na, there was no difference in the concentration of soil organic C, N and Ca along the eutrophication gradient. However, there was a gradient of organic C, N and Ca accumulation corresponding to the area of enhanced peat accretion. The highest rates occurred 1.6 km south of the Hillsboro canal (212 ± 5 g organic C/m²/yr, 14.1 ± 0.4 g N/m²/yr, 22.1 ± 5.2 g Ca/m²/yr). Accumulation of organic C, N and Ca at distances of 7.1–10.7 km downstream averaged 87 ± 11, 6.3 ± 0.7 and 6.5 ± 0.9 g/m²/yr, respectively. The areal extent of enhanced peat accretion and organic C, N, Ca and Na accumulation encompasses approximately 7700 ha of the northern part of WCA 2A. The area of enhanced P accumulation is larger, covering 11,500 ha or 26% of the total area of WCA 2A. The 11,500 ha area has functioned as a sink for P for the past 25–30 yr removing 74% (49.3 MT/yr) of the 67 MT/yr that enters via agricultural drainage and rainfall. Moreover, P accumulation along the gradient was related to mean (1989–1990) surface water P concentration, decreasing as surface water P decreases. These findings suggest that P accumulation is dependent on the P concentration in the water column and that decreasing P loadings per unit area result in less P storage per unit area. The potential longterm equilibrium of the 11,500 ha area as a sink for P is based on a mean annual loading of 67 metric tons P/yr. Input rates exceeding this loading rate could result in an expansion of the 11,500 ha area until a new equilibrium size is reached.     

Fermentative production of acetic acid from various pure and natural cellulosic materials by Clostridium lentocellum SG6

Clostridium lentocellum SG6 fermented various pure crystalline cellulosic materials efficiently with maximum acetic acid yield (gram acetic acid/gram substrate) of 0.67, at low substrate (8 g l⁻¹) concentration. The strain grew poorly on crude biopolymers but fermented them easily after alkali treatment, when grown with 8 g substrate l⁻¹ concentration of alkali-extracted cotton straw (AECS), paddy straw (AEPS) and sorghum stover (AESS) etc. The acetic acid to substrate (A/S) ratios were similar to those obtained with pure cellulosic materials. An increase in substrate concentration led to a decreased A/S ratio and a decreased percentage of substrate degraded. At high substrate concentration of 75 g filter paper l⁻¹, the strain SG6 converted 63.2 g filter paper into 31.28 g acetic acid l⁻¹. At 100 g l⁻¹ concentrations, AECS and AEPS served as the best substrates for acetic acid production when compared with other biopolymers. A maximum amount of 30.98 and 30.86 g acetic acid was produced from 70.6 g AEPS and 70.1 g AESS l⁻¹ of medium by strain SG6, respectively. Acetic acid production of 0.67 g g⁻¹ pure cellulose (Whatman No. 1 filter paper), 0.63 g g⁻¹ of alkali-treated cotton straw (AECS) are the highest among the cellulolytic bacteria reported so far in mono culture fermentations with pure and native cellulosic materials. This revised version was published online in July 2006 with corrections to the Cover Date.     

A case of unusually high oxygen demand in a eutrophic lake

An unusually high hypolimnetic water column BOD (WCBOD), roughly 40 times higher than the sediment oxygen demand (SOD), was observed in a small eutrophic lake and an adjoining lagoon. The mean 5-day WCBOD during thermal stratification in the lake was 29 and 49 g/m² at 10 and 20 °C, respectively, while in the lagoon it was even higher (47 and 87 g/m² at 10 and 20 °C, respectively). The soluble fraction comprised about two-thirds of the WCBOD. WCBOD in the lake was much less during the unstratified period (5-day = 5 g/m²). The SOD rates at two depths in both the lake (0.31 and 0.2 g/m²-d) and lagoon (0.41 and 0.28 g/m²-d) were not unusually high. The ultimate whole BOD (UWCBOD + USOD) was approximately 96 g/m² in the lake and 136 g/m² in the lagoon and UWCBOD formed over 90% of the ultimate whole BOD in both water bodies. A possible cause for these abnormally high WCBODs, in addition to the normal autochthonous production, is an allochthonous source from loosely aggregated and flocculant mats of the bog moss, Sphagnum, which surrounds the lake-lagoon system. Storm water per se was clearly insignificant, but would have contributed indirectly through nutrients for autochthonous production. Such high short-term BOD rates may greatly over-estimate the demand to be satisfied by continuous aeration.     

Enhancement of tryptophan production by Escherichia coli as an application of genetic engineering

Summary A feedback resistant trp operon plasmid that transformed a multiple mutant (trpR tnaA) of Escherichia coli was found to enhance remarkably the production of tryptophan in a bench-scale fermentor. 5.5 g of tryptophan was accumulated per litre of culture medium at 24th hr in batch. The productivity was 0.229 g/l/hr. This productivity is the highest among those ever reported by other workers. The recombinant plasmid (Tc^r Trp⁺ I^-) used was completely stable in each run when tetracycline was added by 10 g/l into the medium.     

The growth of four species of Azolla as affected by temperature

The growth of 22 strains of Azolla pinnata R. Br., 3 strains of A. filiculoides Lam. and one strain each of A. mexicana Presl and A. caroliniana Willd. was tested separately in liquid culture media kept in controlled, artificial light (30 klux) growth cabinets. Three temperature levels were used: 33°C (37/29°C day/night), 29°C (33/25°C) and 22°C (26/18°C)/ Photoperiod was 12 h a day.For most A. pinnata strains (except three) and an A. mexicana strain the maximum weekly relative growth rate was higher at 33°C than at 22°C, but not for A. filiculoides and A. caroliniana. The highest value of maximum relative growth rate corresponded to 1.9 doubling days and in most strains this occurred in the first week. As the plants grew, the growth rate slowed down more severely at higher temperatures. The maximum biomass was higher at 22°C than at 33°C in all strains. At 22°C, it took 30–50 days to attain maximum biomass and the highest value was 14 g N m^?2 or 320 g dry m^?2 by A. caroliniana, followed by 12 g N m^?2 or 290 g dry wt. m^?2 by one strain of A. filiculoides. At 29°C, the maximum biomass was attained in 20–35 days. The highest value was 6.3 g N m^?2 or 154 g dry wt. m^?2 by A. caroliniana. At 33°C, most A. pinnata strains gave a maximum biomass of less than 4 g N m^?2 after 13–23 days, while some strains grew up to 30 days, resulting in a higher maximum biomass. The highest maximum biomass at 33°C was 5.5 g N m^?2 or 140 g m^?2 dry wt. by A. pinnata from Cheng Mai while the maximum biomass of A. filiculoides and A. caroliniana was much less. Azolla filiculoides requires lower temperature than other species for its growth. Azolla pinnata has the best tolerance to high temperatures among the four species. Azolla mexicana could not be discriminated from A. pinnata in its response to temperature. Azolla caroliniana may keep an intermediate position between A. filiculoides and A. pinnata in temperature response.The formation of ammonia in the medium was examined and it occurred mostly under stationary growth conditions, but, at 33°C, some strains of A. pinnata and A. mexicana released or formed ammonia at 0.3–0.8 μg N ml^?1 per week during their initial exponential growth stage.     

A general,fast, and sensitive micromethod for DNA determination: Application to rat and mouse liver,rat hepatoma,human leukocytes,chicken fibroblasts,and yeast cells

A fluorometric method using 3,5-diaminobenzoic acid for DNA determination in tissues, cultured cells, nucleated blood cells, and yeast cells is described. The method is general, simple, and rapid, and does not require prior DNA extraction, since tissue is directly solubilized in Triton X-100 and ammonia. The procedure is highly sensitive, and is able to measure rather accurately as little as 10 ng of DNA. It is applicable to all types of DNA structure. The DNA content determined in various tissues and cells was: 2.50 mg/g fresh rat liver, 3.32 mg/g rat diethylnitrosamine-induced hepatoma, 2.49 mg/g fresh mouse liver, 8.76 μg/10⁶ human leukocytes, 3.37 μg/10⁶ chicken fibroblasts, 2.97 μg/10⁸ haploid yeast cells, and 2.84 μg/10⁸ haploid yeast protoplasts.     

ROOT PRODUCTION IN FOUR COMMUNITIES IN THE GREAT DISMAL SWAMP

A sequential coring approach was used to measure root biomass and production over 1 year in four different communities within the Great Dismal Swamp. A second method, an implanted bag technique, was also used to measure root production, and values were generally lower using this technique. On all sites, fine roots were the most dynamic root component. Both biomass (1,887 g/m²) and production (354–989 g m ² yr^-1) were highest on the mixed hardwood site, the least flooded site, and second highest on the cedar site, the site with the longest duration of soil saturation (1,033 g/m² and 274–366 g m^-2 yr^-1). The maple-gum (696 g/m² and 59–91 g m^-2 yr^-1) and cypress (824 g/m² and 68–308 g m^-2 yr^-1) sites had similarly low amounts of biomass and rates of production. Environmental parameters that influenced production include frequency and duration of flooding, and soil type. Peaks in belowground production were observed on the most productive sites (mixed hardwood and cedar) in summer and late fall-winter; the other two sites exhibited little seasonal variability. The least flooded stand appears to allocate a greater percentage of net primary production belowground than the more extensively flooded stands. The ratio of above- and belowground allocation appears to change in response to a flooding gradient. This has major implications for ecosystem functions as carbon allocation patterns determine the array of litter types generated (leaves vs. roots) which affect decomposition rates and nutrient availability.     

Synthesis and biological activity of [L-3,4-dehydroproline3]-tuftsin

A 3,4-dehydroproline analogue of tuftsin (L-Thr-L-Lys-L-Pro-L-Arg) was prepared by the solid phase synthetic method. Following reversed-phase high performance liquid chromatography (HPLC) purification, the analogue was compared to tuftsin for its ability to enhance the chemotactic, bactericidal and phagocytic activities of polymorphonuclear leukocytes (PMN). Both tuftsin and [Δ³-pro³]-tuftsin elicited a similar significant chemotactic effect at a concentration of 10 μg/ml. A slight suppression of the chemotactic activity was observed with tuftsin at 10^?3 μg/ml and with [Δ³-pro³]-tuftsin at concentrations of 10^?3, 10^?2 (significant) and 10^?1 μg/ml. Although similar bactericidal activities were observed for both peptides, PMN exposed to [Δ³-pro³]-tuftsin exhibited increased phagocytic indicies 2–4 times that of tuftsin-treated PMN at concentrations of 0.4, 0.6 and 1.0 μg/ml.     

Production and ecology of eelgrass (Zostera marinal L.) in the Grevelingen estuary,the Netherlands,before and after the closure

The Grevelingen estuary was cut off from the North Sea and from the influences of the river Rhine by a dam in 1971, and became a stagnant salt-water lake. Production and ecology ofZostera marina L. were studied in 1968 and in 1973–1975, both through standing stock estimations, biomass increases in permanent quadrats, and correlation of distribution patterns with ecological factors. After the closure of the estuary the intertidal eelgrass population extended downwards to 5 m below lake level, probably owing to the increased transparency of the water; the area occupied, and the density of the eelgrass beds increased strongly. Eelgrass annual overground production, based on doubled maximum standing crop values in July–August, was estimated at 50 g C/m² in 1968, 121 g C/m² in 1973 and 91 g C/m² in 1975 inZostera beds, and 4 g C/m² in 1968, 18 g C/m² in 1973 and 23 g C/m² in 1975 for the entire Grevelingen area. A minimum estimate of net production inZostera beds at a depth of 0.50–0.75 m, based on short term changes in biomass in 2 permanent quadrats in 1974 and 1975, was 40.5 g C/m²/yr for overground parts and 12.7 g C/m²/yr for underground parts. Horizontal distribution of celgrass is not primarily limited by grainsize distribution, but more by exposure to wave action and currents. On account of irradiance reduction light is a limiting factor in the vertical distribution of the eelgrass population in Lake Grevelingen. Communication no. 146 of the Delta Institute for Hydrobiological Research, Yerseke, The Netherlands.     

Flow injection chemiluminescence determination of 2‐methoxyestradiol based on inhibition of luminol‐potassium ferricyanide reaction

A novel flow‐injection chemiluminescence (FI‐CL) method is described for the determination of 2‐methoxyestradiol (2‐ME). The method is based on the inhibitory effect of 2‐ME on the CL reaction of luminol and potassium ferricyanide in alkaline solution. Under optimal conditions, net CL intensity was proportional to 2‐ME concentration in synthetic and mouse plasma samples. Corresponding linear regression equations were 8.0 x 10^‐9‐1.0 x 10^‐7g/mL for synthetic samples and 2.0 x 10^‐9‐1.0 x 10^‐7g/mL for plasma samples. Detection limit for synthetic samples and limits for quantification of plasma samples were 8.4 x 10^‐10g/mL (3σ) for synthetic samples and 4.0 x 10^‐9g/mL for mouse samples. A complete analysis was performed for 60 s, including washing and sampling, resulting in a throughput of ≈ 60/h. The proposed method was applied for the determination of 2‐ME in synthetic and mouse plasma samples. Percentage recoveries were 101.0‐102.8% and 98.0‐105.0%, respectively. A possible mechanism responsible for CL reaction is proposed. Copyright © 2012 John Wiley & Sons, Ltd.     

离子转运蛋白调控钝齿棒杆菌离子和pH稳态促进L-精氨酸合成

L-精氨酸是一种半必需氨基酸,广泛应用于食品、制药、饲料等行业。【目的】当前对L-精氨酸生产菌株的研究,极少涉及离子转运领域。在本研究中,发现在发酵时适量添加外源K~+有利于促进钝齿棒杆菌(Corynebacterium crenatum) SYPA5-5合成L-精氨酸。【方法】在C. crenatum SYPA5-5发酵培养基外源添加0.5 g/L和2.5 g/L的K_3PO_4,取对数期发酵样品进行转录组数据分析,挖掘出K~+转运相关的阳离子转运ATP酶CTAP1以及单价阳离子/H~+逆转运蛋白Mrp1A,研究其在C. crenatum SYPA5-5快速合成L-精氨酸阶段,对菌株生长及L-精氨酸合成的影响。【结果】对基因ctap1和mrp1分别进行敲除和过表达,深入研究突变株对L-精氨酸合成的影响。研究发现同时过表达离子转运蛋白CTAP1和Mrp1A更有利于胞内离子、pH稳态和渗透压调节,最终提高L-精氨酸的产量。在补料分批发酵中分别过表达Mrp1A、CTAP1以及同时过表达Mrp1A和CTAP1的菌株L-精氨酸产量分别达到61.4 g/L、63.9 g/L和65.3 g/L,产率分别为0.383 g/g、0.392 g/g和0.395 g/g,比C. crenatum SYPA5-5分别提高了34.9%、38.0%和39.1%。【结论】CTAP1是特异性的K~+转运ATP酶,可以将培养基中的K~+运输到胞内。同时Mrp1A可将胞内K~+和Na~+等单价阳离子运输到胞外,将胞外H~+运输至胞内,中和胞内L-精氨酸所导致的碱性环境,从而维持胞内pH稳定。CTAP1和Mrp1A的研究为解析离子转运机制和L-精氨酸合成之间的联系奠定了基础。