尿素与磷酸二氢钾配施对板栗光合特性及生长结实的影响

肥料配施可提高肥料利用效率,改善树体营养结构,起到平衡施肥的作用.选取7年生板栗树为试材,采用树干注射的方法,研究尿素和磷酸二氢钾不同配施处理对板栗光合特性及生长结实的影响.结果表明: 尿素和磷酸二氢钾配施具有明显的正向协同效应,二者配施较单一施肥可显著提高板栗光合能力及产量和品质.单施(NH₂)₂CO可降低叶绿素含量,单施KH₂PO₄可增加叶绿素含量,而二者配施使叶绿素含量显著增加.4种配施处理均可提高叶片及枝条的N、P、K含量,其中0.3%(NH₂)₂CO+0.3%KH₂PO₄处理效果最好.不同施肥处理均可改善光合参数,但仍以配施处理为好,其中0.3%(NH₂)₂CO+0.3%KH₂PO₄处理可显著提高光合速率、最大净光合、表观量子效率、羧化效率、瞬时水分利用效率、氮素利用效率.配施可同时促进枝条加长和加粗生长,并提高混合芽数量,而单施(NH₂)₂CO仅能促进加长生长,对提高混合芽数量效果不显著.配施对提高坚果产量和质量效果优于单一施肥,在0.3%(NH₂)₂CO+0.3%KH₂PO₄处理下坚果产量、单粒质量和总糖含量等关键指标较对照分别提高68.2%、25.5%和14.9%.     

Optimization of culture conditions for production of cis-epoxysuccinic acid hydrolase using response surface methodology

Response surface methodology, which allows for rapid identification of important factors and optimization of them to enhance enzyme production, was employed here to optimize culture conditions for the production of cis-epoxysuccinic acid hydrolase from Bordetella sp. strain 1–3. In the first step, a Plackett–Burman design was used to evaluate the effects of nine variables (yeast extract, cis-epoxysuccinic acid, KH₂PO₄, K₂HPO₄ · 3H₂O, MgSO₄ · 7H₂O, trace minerals solution, culture volume, initial pH and incubation time) on the enzyme production. Yeast extract, cis-epoxysuccinic acid and KH₂PO₄ had significant influences on cis-epoxysuccinic acid hydrolase production and their concentrations were further optimized using central composite design and response surface analysis. A combination of adjusting the concentration of yeast extract to 7.8 g/l, cis-epoxysuccinic acid to 9.8 g/l, and KH₂PO₄ to 1.12 g/l would favor maximum cis-epoxysuccinic acid hydrolase production. An enhancement of cis-epoxysuccinic acid hydrolase production from 5.6 U/ml to 9.27 U/ml was gained after optimization.     

响应面法优化荷叶离褶伞菌丝体产漆酶条件

为了对荷叶离褶伞产漆酶条件进行优化,在单因素实验基础上,通过最陡爬坡实验(PB)对培养基8因素进行筛选,获得影响产漆酶的3个显著性因素:葡萄糖,pH和KH₂PO₄;通过中心组合(CCD)设计及响应面分析确定了最优发酵条件:葡萄糖20.09g/L,酪蛋白1.5g/L,酵母提取物1.5g/L,MgSO₄ 3g/L,CuSO₄ 3.75mg/L,KH₂PO₄ 3.97g/L,pH 4.98,VB₁ 0.1g/L,愈创木酚12mg/L,该条件下,漆酶酶活为829.83U/mL,较未优化对照提高46.6%.     

黑曲霉生产β-葡萄糖苷酶发酵条件的研究产

经多项式回归分析,研究了不同浓度N源、C源、无机盐等对酶产量的影响,确定出最佳培养基配方为:麸皮4.9%,(NH₄)_2>SO₄0.4%,KH₂PO₄0.29%,CaCl₂0.05%,MgSO₄·7H₂O0.04%,FeSO₄·7H₂O 5mg·L^-1,ZnCl₂1.4mg·L^-1,0.2%油酸钠。并对培养温度、时间、培养基初始pH、通气量、接种量、接种方式等培养条件进行优化,使黑曲霉生产β葡萄糖苷酶的产量由17U·ml^-1增至21.3U·ml^-1.     

The optimization of manganese dioxide bioleaching media by fractional factorial experiments

The microbial leaching of manganiferous minerals originating from Greece and Italy has been the subject of EU research for about 3 years, in order to develop an economical process of biotreatment.

A heterotrophic mixed culture, selected from natural water, was found to be capable of reducing the MnO₂ in microaerobic conditions. The first culture medium, formulated in a preliminary way, was employed in the selection and cultivation of the microbial strains in the first steps of this study. For this reason, an optimization study of the culture media was carried out by means of fractional and full factorial experiments in order to establish the correct medium composition for the biotreatment. The bioleaching tests were carried out in shake flasks using a low grade Italian mineral. The factors analysed were the concentrations of minerals, molasses, NH₄NO₃, KH₂PO₄, MgSO₄, yeast extract and NaHCO₃. The manganese extraction yield was the response of the process. A reduction of the salt concentrations present in the media was found to be possible with a significant reduction in cost. The experimental results show the potential application of the mixed culture for the bioleaching of manganiferous minerals not treatable by means of conventional processes.     

响应面分析法优化黑根霉胞外多糖发酵工艺

实验以商品化的马铃薯葡萄糖液体培养基为基础培养基,以胞外粗多糖产量为考察指标,运用响应面分析法考察玉米浆浓度、KH₂PO₄浓度和发酵时间3个因素对胞外多糖发酵产量的影响,以获得黑根霉胞外多糖发酵最优工艺,建立高产、稳定、可控的胞外多糖发酵生产工艺技术方案。经响应面分析,各因素按照对响应值的影响顺序为：玉米浆浓度>发酵时间> KH₂PO₄浓度,且玉米浆浓度、发酵时间对胞外多糖产量的影响极显著,KH₂PO₄浓度对胞外多糖产量的影响不显著。胞外多糖发酵最优工艺为：玉米浆3.2mg/mL、KH₂PO₄ 1.5mg/mL和发酵时间132h,在此条件下胞外多糖的最大预测产量为0.824mg/mL。实验重复性好,是一个高产、稳定、可控的胞外多糖发酵生产工艺技术方案,可以指导黑根霉胞外多糖发酵。     

肺形侧耳发酵培养基优化及多糖提取工艺

通过碳源、氮源单因素实验和正交实验,对野生肺形侧耳进行发酵培养基优化。选取浸提时间、浸提温度及液料比3个因素,以胞内粗多糖提取率为指标,采用正交实验设计确定菌丝体胞内多糖提取的最佳工艺。结果表明,适宜肺形侧耳深层发酵的培养基为蔗糖1.5%,麸皮5%,蛋白胨0.6%,KH₂PO₄ 0.15%,MgSO₄ 0.75%,VB₁ 0.01%。胞内多糖提取的最佳工艺为浸提时间2h,液料比50:1,浸提温度90℃,此条件下多糖提取率为34.35%。     

Medium improvement by orthogonal array designs for cholesterol oxidase production by Rhodococcus equi No. 23

Medium improvement for the production of cholesterol oxidase (CO, EC 1.1.3.6) by Rhodococcus equi No. 23 was investigated using an orthogonal array design in two steps. Results revealed that yeast extract, Tween 80 and zinc sulphate had positive effects on CO production, but magnesium sulphate had an inhibitory effect. In addition, interaction between cholesterol and sodium chloride also had a significant effect on enzyme production. The improved medium consisted of 2·0 g/litre cholesterol, 8·0 g/litre yeast extract, 1·0 g/litre NH₄Cl, 1·0 g/litre NaCl, 0·50 g/litre KH₂PO₄, 0·25 g/litre Na₂HPO₄, 0·10 g/litre -valine, 0·15 g/litre -tyrosine, 0·15 g/litre MgSO₄·7H₂O, 0·01 g/litre ZnSO₄·7H₂O, 0·10 g/litre FeSO₄·7H₂O and 4·0 ml/litre Tween 80. CO production at 60 h (about 0·24 units/ml) was about four-fold greater than with the control medium.     

落叶松锈迷孔菌产多糖液体培养基的优化及其体外抗氧化活性

本研究通过响应面设计优化了落叶松锈迷孔菌产胞外多糖的液体培养基,并对其体外抗氧化活性进行了分析。结果显示,以落叶松锈迷孔菌胞外多糖产量为响应值,采用Plackett-Burman设计、最陡爬坡设计和Box-Behnken设计建立数学模型,获得落叶松锈迷孔菌产胞外多糖的最适培养基为：去皮马铃薯415.70g/L、蛋白胨10.80g/L、葡萄糖20.0g/L、吐温80 7.27mL/L、KH₂PO₄ 1.0g/L、MgSO₄?7H₂O 0.5g/L、CaCO₃ 0.57g/L、FeSO₄?7H₂O 0.3g/L、ZnSO₄?7H₂O 0.3g/L、维生素B₁ 0.01g/L、芦丁6.09g/L。在此优化条件下得到的多糖产量为3.07g/L,与理论值3.06g/L相近,相比于优化前提高了2.87倍,说明响应面法对于优化落叶松锈迷孔菌的液体培养基以提高多糖产量行之有效。此外,经过醇沉、透析和去蛋白等获得初步纯化的多糖可有效清除羟自由基、超氧阴离子、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和2,2?-连氮-双(3-乙基苯并噻唑-6-磺酸)[2,2?-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid),ABTS]自由基。     

Optimization of media for β-fructofuranosidase production by Aspergillus niger in submerged and solid state fermentation

The kinetics of β-fructofuranosidase (Ffase) production by Aspergillus niger in submerged (SmF) and solid-state fermentation (SSF) systems was investigated. The maximum productivity of Ffase (81.8 U/l per h) was obtained in SSF for 72 h while it was 18.3 U/l per h in SmF for 120 h. The productivity of extra cellular Ffase produced in SSF was 5-fold higher than in SmF. Optimization of fermentation medium for Ffase production was carried out using De Meo's fractional factorial design with seven components such as (NH₄)₂SO₄, KH₂PO₄, FeSO₄, MgSO₄ · 7H₂O, sucrose, urea and yeast extract. The media designed for SmF after two steps of optimization supported the growth of A. niger and higher productivity of Ffase (58.3 U/l per h) than with the medium before optimization. The optimized medium of SmF when used in SSF, did not improve the Ffase productivity and therefore medium for SSF was optimized independent of SmF. After two optimization steps, the media was defined for SSF which supported the growth and high level of Ffase productivity (149.1 U/l per h) in SSF compared to the medium before optimization (81.8 U/l per h) and optimized medium for SmF (58.3 U/l per h). Our results suggested that the optimized media for SmF and SSF for the production of Ffase have to be different.     

Platinum(II) complexes of 4-methoxy- and 4-chlorobenzoic acid hydrazides. Synthesis, characterization, and cytotoxic effect

The complexes [Pt(NH₃)(pmbah)Cl₂], [Pt(NH₃)(pcbah)Cl₂], [Pt(pmbah₂X₂] and [Pt(pcbah)₂X₂] (pmbah = 4-methoxybenzoicacid hydrazide, pcbah = 4-chlorobenzoic acid hydrazide; X = Cl, Br, I) have been synthesized and characterized by elemental analysis, electric conductivity, ¹H NMR, IR, and electronic spectra. A cis-square planar structure with hydrazide ligands coordinated via the NH₂ groups has been proposed for these compounds. The complexes, but not the free ligands, have shown a strong growth inhibitory effect in Friend leukemia cells in vitro, most of which are more active than cisplatin.     

Practical Methods for the Control of Hydrogenion Concentration in Staining Procedures—The Use of “Buffers”

Data are given for the simple preparation of dry chemical mixtures to be used in the easy preparation of buffer solutions for use in the control of stain procedures. Two of these mixtures of special value in practical staining operations are the following: 4.539 g. KH₂PO₄, 5.940 g. Na₂.HPO₄.2H₂O. (pH=6.8); 7.262 g. KH₂PO₄, 2.376 g Na₂HPO₄.2H₂O (pH = 7.4). Hydrogen-ion control has been found to be essential to consistent results with the compound blood and tissue stains such as Wright's and Giemsa preparations. The more general use of a pre-determined pH in staining procedures is indicated; several possible methods of application are suggested.     

一株病原拮抗野生菌株的筛选、鉴定及其发酵工艺优化

为获得广谱抗菌功能野生菌株并提高其发酵产物中抗菌物质的含量。采用管碟法和菌丝生长速率法筛选功能菌株,ITS序列分析鉴定功能菌株,通过响应面法和正交设计优化发酵生产抗菌物质的工艺。筛选到一株强效、广谱抗菌功能菌株,鉴定为Cerrena sp.,其发酵产物对金黄色葡萄球菌、大肠杆菌、白色念珠菌、枯草芽孢杆菌和水稻纹枯病菌有显著拮抗作用。该菌株的摇甁发酵配方及培养条件为：马铃薯13.99 g/L,蔗糖 41.58 g/L,VB1 0.027 g/L,麸皮7 g/L,KH₂PO₄ 2 g/L,MgSO₄·7H₂O 2 g/L;摇床温度28 ℃、发酵周期10 d、种龄4 d、接种量8%、初始pH为5.0、装液量110 ml/250 ml。该菌株有明显抑菌活性,发酵工艺优化后抗菌活性提高了30.37%,为该菌株今后的应用、抗菌剂的分离提纯和产业化提供了实验依据。     

New extracellular resistance mechanism for cisplatin

The HSQC NMR spectrum of ¹⁵N-cisplatin in cell growth media shows resonances corresponding to the monocarbonato complex, cis-[Pt(NH₃)₂(CO₃)Cl]⁻, 4, and the dicarbonato complex, cis-[Pt(NH₃)₂(CO₃)₂]⁻², 5, in addition to cisplatin itself, cis-[Pt(NH₃)₂Cl₂], 1. The presence of Jurkat cells reduces the amount of detectable carbonato species by (2.8 ± 0.7) fmol per cell and has little effect on species 1. Jurkat cells made resistant to cisplatin reduce the amount of detectable carbonato species by (7.9 ± 5.6) fmol per cell and also reduce the amount of 1 by (3.4 ± 0.9) fmol per cell. The amount of detectable carbonato species is also reduced by addition of the drug to medium that has previously been in contact with normal Jurkat cells (cells removed); the reduction is greater when drug is added to medium previously in contact with resistant Jurkat cells (cells removed). This shows that the platinum species are modified by a cell-produced substance that is released to the medium. Since the modified species have been shown not to enter or bind to cells, and since resistant cells modify more than non-resistant cells, the modification constitutes a new extracellular mechanism for cisplatin resistance which merits further attention.     

六妹羊肚菌工厂化制备液体菌种繁育条件的优化

为适合大规模工厂化制备羊肚菌液体菌种,以提高菌丝体生物量为目的,控制菌丝球直径,使用六妹羊肚菌Morchella sextelata,采用单因素、Plackett-Burman、响应面Box-Behnken实验摇瓶培养优化液体菌种制备条件,发酵罐扩大培养,具体参数为玉米淀粉2.5%、麸皮6%、酵母粉0.4%、葡萄糖2.5%、蛋白胨0.25%、磷酸二氢钾0.1%、硫酸镁0.05%、接种量10%、pH 5.5、转速200-500r/min、温度25℃、培养1-3d,在该条件下羊肚菌菌丝体干重可达16mg/mL,菌丝球直径降至1.5mm以下,密度达到2 967个/mL,从母种到栽培种仅需13-19d。本研究结果为羊肚菌工厂化液体菌种制备和栽培提供了理论基础。     

K88大肠杆菌菌毛蛋白发酵工艺条件优化研究

分别采用LB培养基、牛肉膏蛋白胨培养基、强化营养培养基、玉米浆培养基对大肠杆菌K88进行发酵培养,选出最适于大肠杆菌K88生长的玉米浆培养基;采用正交实验对玉米浆培养基的C／N、K2HPO4／KH2PO4、Mg^2＋的配比进行优化,筛选最适于大肠杆菌K88生长的营养配比;研究生长曲线、接种量以及菌体和菌毛生产量的相关性,根据实验结果优化发酵培养条件,确定菌种的最佳发酵工艺,以收获最多的K88菌毛蛋白。研究表明,K88大肠杆菌在玉米浆培养基C／N、K2HP04／KH2PO4的配比分别为5／11、1／1,Mg^2＋为0．1g/mL,pH值为7．2,转速为200r／min,接种量为4．5％的条件下发酵26个小时,菌体和菌毛生产量均达到高峰,同时得出菌毛蛋白产生量和菌体量成正相关。     

Protein-based complex medium design for recombinant serine alkaline protease production

This work reports on the design of a complex medium based on simple and complex carbon sources, i.e. glucose, sucrose, molasses, and defatted-soybean, and simple and complex nitrogen sources, i.e. (NH₄)₂HPO₄, casein, and defatted-soybean, for serine alkaline protease (SAP) production by recombinant Bacillus subtilis carrying pHV1431::subC gene. SAP activity was obtained as 3050 U cm⁻³ with the initial defatted-soybean concentration C_soybean^o=20 kg m⁻³ and initial glucose concentration C_G^o=8 kg m⁻³; whereas, addition of the inorganic nitrogen source (NH₄)₂HPO₄ decreased SAP production considerably. Further increase in SAP production (3850 U cm⁻³) was obtained when sucrose was replaced with glucose at C_sucrose^o=15 kg m⁻³ and C_soybean^o=20 kg m⁻³. Nevertheless, when molasses was replaced with sucrose, the maximum activity was obtained with molasses having 10 kg m⁻³ initial sucrose concentration and C_soybean^o=15 kg m⁻³as 2130 U cm⁻³; moreover, when casein was replaced with defatted-soybean SAP production decreased considerably (ca. 250 U cm⁻³). Thereafter, the effects of inorganic ionic compounds were investigated; and except phosphate, inorganic compounds supplied from defatted-soybean were found to be sufficient for the bioprocess. The highest SAP activity was obtained as 5350 U cm⁻³ in the medium that contained (kg m⁻³): C_soybean^o=20, C_sucrose^o=15, C_Na2HPO4^o=0.021, and C_NaH2PO4^o=2.82, that was 6.5-fold higher than that of the SAP produced in the defined medium. By using the designed complex medium, oxygen transfer characteristics of the bioprocess were investigated; and, Damköhler number that is the oxygen transfer limitation increases with the cultivation time until t=14 h; and, at t>20 h both mass transfer and biochemical reaction resistances were effective. Overall oxygen transfer coefficient varied between 0.010 and 0.044 s⁻¹; volumetric oxygen uptake rate varied between 0.001 and 0.006 mol m⁻³ s⁻¹; and specific oxygen uptake rate varied between 0.0001 and 0.0022 mol kg⁻¹ DW s⁻¹ throughout the bioprocess.     

野生四川灵芝的生物学特性和抗氧化活性

灵芝是我国一类传统药用真菌的统称,具有极高的药用价值和经济价值.为了充分保护和利用该类野生药用真菌资源,本研究对采自广西壮族自治区南宁市的一株野生灵芝进行了菌株分离纯化培养,通过基于内转录间隔区(internal transcribed spacer,ITS)序列的分子生物学分析鉴定为四川灵芝Ganoderma sic...     

高温下喷施水杨酸和磷酸二氢钾对中稻生理特征和产量的影响

为探寻减轻水稻抽穗开花期高温热害的技术途径,以3个籼型杂交稻品种为材料,于2017—2018年在江西省吉安县、余干县、南昌县进行田间试验.在抽穗开花期自然高温下,设置叶面喷施5个不同浓度的水杨酸(SA)处理(SA₁～SA₅分别为100、500、1000、1500、2000 μmol·L^-1)和5个不同浓度的磷酸二氢钾(KH₂PO₄)处理(K₁～K₅分别为7.35、14.70、22.05、29.40、36.75 mmol·L^-1),并以叶面喷施蒸馏水为对照(CK),分析中稻生理特征和产量.结果表明: 与对照相比,SA处理和KH₂PO₄处理分别降低了剑叶丙二醛含量,提高了叶绿素、可溶性糖、可溶性蛋白、脯氨酸含量和超氧化物歧化酶、过氧化物酶活性,其中SA₂和K₃处理效果最好.SA₂、SA₃和K₃、K₄处理提高了水稻穗粒数、结实率和产量,其中SA₂和K₃处理效果显著,与对照相比,SA₂处理分别使穗粒数、结实率和产量增加了7.0%、4.0%和11.9%,K₃处理增加了3.9%、4.7%和6.6%.抽穗开花期高温下,采取叶面喷施500 μmol·L^-1 SA或22.05 mmol·L^-1 KH₂PO₄的技术措施可显著提高中稻产量.     

Synthesis of tripeptide RGD amide by a combination of chemical and enzymatic methods

The tripeptide Bz-Arg-Gly-Asp(NH₂)OH was synthesized by a combination of chemical and enzymatic methods in this study. Firstly, Gly-Asp-(NH₂)₂ was synthesized by a novel chemical method in three steps including chloroacetylation of l-aspartic acid, esterification of chloroacetyl l-aspartic acid and ammonolysis of chloroacetyl l-aspartic acid diethyl ester. Secondly, the linkage of the third amino acid (Bz-Arg-OEt) to Gly-Asp-(NH₂)₂ was completed by enzymatic method under kinetic control condition. An industrial alkaline protease alcalase was used in water–organic cosolvents systems. The synthesis reaction conditions were optimized by examining the effects of several factors including water content, temperature, pH and reaction time on the yield of the synthesis product Bz-Arg-Gly-Asp(NH₂)OH. The optimum conditions are pH 8.0, 35 °C, in ethanol/Tris–HCl buffer system (85:15, v/v), 8 h with the tripeptide yield of 73.6%.