全文获取类型
收费全文 | 392篇 |
免费 | 24篇 |
国内免费 | 31篇 |
专业分类
化学 | 412篇 |
综合类 | 14篇 |
数学 | 1篇 |
物理学 | 20篇 |
出版年
2023年 | 3篇 |
2022年 | 8篇 |
2021年 | 26篇 |
2020年 | 8篇 |
2019年 | 13篇 |
2018年 | 5篇 |
2017年 | 14篇 |
2016年 | 10篇 |
2015年 | 16篇 |
2014年 | 8篇 |
2013年 | 13篇 |
2012年 | 36篇 |
2011年 | 18篇 |
2010年 | 16篇 |
2009年 | 15篇 |
2008年 | 22篇 |
2007年 | 21篇 |
2006年 | 14篇 |
2005年 | 20篇 |
2004年 | 15篇 |
2003年 | 21篇 |
2002年 | 16篇 |
2001年 | 14篇 |
2000年 | 13篇 |
1999年 | 11篇 |
1998年 | 9篇 |
1997年 | 16篇 |
1996年 | 8篇 |
1995年 | 4篇 |
1994年 | 5篇 |
1993年 | 3篇 |
1992年 | 5篇 |
1991年 | 3篇 |
1990年 | 3篇 |
1989年 | 2篇 |
1987年 | 1篇 |
1986年 | 2篇 |
1984年 | 3篇 |
1983年 | 1篇 |
1982年 | 1篇 |
1981年 | 2篇 |
1980年 | 1篇 |
1977年 | 1篇 |
1975年 | 1篇 |
排序方式: 共有447条查询结果,搜索用时 15 毫秒
441.
We report the first application of hydrophobic interaction between graphene oxide (GO) and negatively charged enzymes to fabricate CE-integrated immobilized enzyme microreactors (IMERs) by a simple and reliable immobilization procedure based on layer by layer assembly. L -lactate dehydrogenase (L -LDH), which is negatively charged during the enzymatic reaction, is selected as the model enzyme. Various spectroscopic techniques, including SEM, FTIR, and UV-vis are used to characterize the fabricated CE-IMERs, demonstrating the successful immobilization of enzymes on the negatively charged GO layer in the capillary surface. The IMER exhibits excellent repeatability with RSDs of inter-day and batch-to-batch less than 3.49 and 6.37%, respectively, and the activity of immobilized enzymes remains about 90% after five-day usage. The measured Km values of pyruvate and NADH of the immobilized L -LDH are in good agreement with those obtained by free enzymes. The results demonstrate that the hydrophobic interactions and/or π-π stacking is significant between the GO backbone and the aromatic residues of L -LDH and favorable to fabrication of CE-integrated IMERs. Finally, the method is successfully applied to the determination of pyruvate in beer samples. 相似文献
442.
L-苯甘氨酸是重要的手性非天然α-氨基酸,可广泛用于合成多种食品添加剂及药物中间体,探索其绿色合成工艺具有重要的意义。 本研究将新型高活性的D-扁桃酸脱氢酶(LhDMDH)和L-亮氨酸脱氢酶(EsLeuDH)偶联,在辅酶内循环的前提下,仅需较低浓度的辅酶即可实现生物催化D-扁桃酸合成L-苯甘氨酸。 通过对加酶量、氧化型烟酰胺腺嘌呤二核苷酸(NAD+)浓度、NH4+浓度和底物浓度等因素进行优化,获得了一个最经济的反应条件:200 mmol/L的D-扁桃酸、6.5 kU/L的加酶量、0.1 mmol/L NAD+、0.5 mol/L NH4+的条件下、30 ℃,反应12 h,在此条件下,产物的得率和对映体过量(e.e.)值分别可达98%和99%以上,具有较大的产业化潜力,为实现L-苯甘氨酸规模化的生物合成奠定了坚实的基础。 相似文献
443.
Improved biosensors for acetaldehyde determination have been developed using a bienzymatic strategy, based on a mediator-modified carbon film electrode and co-immobilisation of NADH oxidase and aldehyde dehydrogenase. Modification of the carbon film electrode with poly(neutral red) mediator resulted in a sensitive, low-cost and reliable NADH detector. Immobilisation of the enzymes was performed using encapsulation in a sol-gel matrix or cross-linking with glutaraldehyde. The bienzymatic biosensors were characterized by studying the influence of pH, applied potential and co-factors. The sol-gel and glutaraldehyde biosensors showed a linear response up to 60 μM and 100 μM, respectively, with detection limits of 2.6 μM and 3.3 μM and sensitivities were 1.7 μA mM−1 and 5.6 μA mM−1. The optimised biosensors showed good stability and good selectivity and have been tested for application for the determination of acetaldehyde in natural samples such as wine. 相似文献
444.
Mammalian mitochondrial dihydrolipoamide dehydrogenase (DLDH, EC 1.8.1.4) catalyzes NAD(+)-dependent oxidation of dihydrolipoamide in vivo and can also act as a diaphorase catalyzing in vitro nicotinamide adenine dinucleotide (reduced form) (NADH)-dependent reduction of electron-accepting molecules such as ubiquinone and nitroblue tetrazolium (NBT). In this paper, we report a gel-based method for histochemical staining and quantification of DLDH diaphorase activity using blue native PAGE (BN-PAGE). Rat brain mitochondrial extracts, used as the source of DLDH, were resolved by nongradient BN-PAGE (9%), which was followed by diaphorase activity staining using NADH as the electron donor and NBT as the electron acceptor. It was shown that activity staining of DLDH diaphorase was both protein amount- and time-dependent. Moreover, this in-gel activity-staining method was demonstrated to be in good agreement with the conventional spectrophotometric method that measures DLDH dehydrogenase activity using dihydrolipoamide as the substrate. The method was applied to determine levels of DLDH diaphorase activity in several rat tissues other than the brain, and the results indicated a similar level of DLDH diaphorase activity for all the tissues examined. Finally, the effects of thiol-reactive reagents such as N-ethylmaleimide (NEM) and nitric oxide donors on DLDH diaphorase activity were evaluated, demonstrating that, with this method, DLDH diaphorase activity can be determined without having to remove these thiol-reactive reagents that may otherwise interfere with spectrophotometric measurement of DLDH dehydrogenase activity. The gel-based method can also be used as a means to isolate mitochondrial DLDH that is to be analyzed by mass spectral techniques in studying DLDH post-translational modifications. 相似文献
445.
446.
Miyuki Masuda Yusuke Motoyama Kenichi Murata Nobuhumi Nakamura Hiroyuki Ohno 《Electroanalysis》2011,23(10):2297-2301
This paper reports a novel mediator for the oxidation of β‐nicotinamide adenine dinucleotide (NAD+/NADH), an electropolymeric film (pAPRu) of [Ru(NH2‐phen)3]2+. A pAPRu‐modified electrode was prepared via electropolymerization and exhibited catalytic activity toward the electrochemical oxidation of NADH due to the imine moieties of pAPRu. The electrochemical oxidation of ethanol was observed using an alcohol dehydrogenase (ADH)‐immobilized electrode. A compartmentless ethanol/O2 biofuel cell composed of an ADH anode and a bilirubin oxidase cathode was constructed. The maximum current density and the maximum power density of the biofuel cell were 190 µA cm?2 and 31 µW cm?2 (at 0.29 V), respectively. 相似文献
447.
以解木糖赖氨酸芽孢杆菌XX-2为出发菌株,110mmol/L L-赖氨酸单盐酸盐为发酵前体,144h发酵后L-2-氨基己二酸浓度达到10.4mmol/L,产率9.5%。以解木糖赖氨酸芽孢杆菌XX-2全细胞为生物催化剂,利用共生的L-赖氨酸 6-脱氢酶和?-1-哌啶啉-6-羧酸脱氢酶催化L-赖氨酸单盐酸盐转化为L-2-氨基己二酸。最优条件为:细胞浓度45g(干重)/L,L-赖氨酸单盐酸盐浓度100mmol/L,pH7.0,温度30℃,反应时间144h。在最优条件下,从100mmol/LL-赖氨酸单盐酸盐产生90mmol/L L-2-氨基己二酸,产率90%。推测了生物催化过程中L-2-氨基己二酸产生的反应机理。 相似文献