微生物发酵法生产S-腺苷甲硫氨酸的研究进展

S-腺苷甲硫氨酸(S-adenosyl-l-methionine, SAM)广泛存在于生物体内,主要参与生物体内的转甲基过程、转硫过程及转氨丙基过程,具有重要的生理功能,其生产备受重视。目前SAM生产的研究主要集中于微生物发酵法,该方法与化学合成法和酶催化法相比,成本较低且更容易实现工业化生产。随着需求量的迅速增加,通过菌种改良提高SAM产量备受关注。当前SAM生产菌种改良的主要策略包括常规育种和代谢工程。本文综述了提高微生物生产SAM能力的近期研究进展并探讨了SAM生产中的瓶颈问题及解决方法,以期为进一步提高SAM产量提供思路。     

Evaluation of total contents of Al,As, Ca,Cd, Fe,K, Mg,Ni, Pb,Zn and their fractions leached to the infusions of different tea samples. A multivariate study

ABSTRACT

Total aluminum, arsenic, calcium, cadmium, iron, lead, potassium, magnesium, nickel and zinc were determined in black tea by electro thermal atomic absorption spectrometry after ultrasonic assisted pseudo-digestion with mixture of acid and oxidant. All the metals were also determined in the tea infusions in order to know the percentage of each element leached into the liquor. A conventional acid digestion on electric hot plate was used to obtain total metals under study for comparative purpose. Analytical results for Al, As, Ca, Cd, Fe, K, Mg, Ni, Pb and Zn obtained by ultrasound assisted pseudodigestion, and conventional wet digestion methods showed a good agreement, thus indicating the possibility of using ultrasonic assisted digestion sample preparation instead of intensive treatments inherent with the acid digestion methods on electric hot plate. The validity and accuracy of both procedures were checked by using certified sample of NIES No. 7 (Tea Leaves). Non significant differences were observed for P>0.05 when comparing the values obtained by both methods (paired t-test).

Principal component analysis (PCA) was applied in order to determine the different amount of metals as main sources of variability in the data sets and to establish the relation between samples (branded and nonbranded tea samples) and micronutrient, trace and toxic metals (variables). Hierarchical cluster analysis (HCA) was used to explore the different branded and nonbranded tea samples grouping according to the essential and toxic metals as additional information to the out put obtained by PCA.     

Aidi injection,a traditional Chinese biomedical preparation for gynecologic tumors: a systematic review and PRISMA-compliant meta-analysis

Aidi injection (ADI), a traditional Chinese biomedical preparation, is a promising adjuvant therapy for gynecologic tumors (GTs), including cervical cancer (CC), endometrial cancer (EC), and ovarian cancer (OC). Although studies have reported positively on ADI therapy, its exact effects and safety in GT patients remain controversial. Therefore, a wide-ranging systematic search of electronic databases was performed for this meta-analysis. Data from 38 trials including 3309 GT patients were analyzed. The results indicated that the combination of conventional treatment and ADI markedly improved the patients’ overall response rate (P<0.00001), disease control rate (P<0.00001), and quality of life (P<0.05) compared with conventional treatment alone. Furthermore, patient immunity was enhanced with combined treatment, as indicated by significantly increased percentages of CD3⁺ (P=0.005) and CD4⁺ (P<0.00001) and increased CD4⁺/CD8⁺ ratio (P=0.001). Most of the adverse events caused by radiochemotherapy such as gastrointestinal issues, leukopenia, thrombocytopenia, and hepatotoxicity, (P<0.05 for all) were significantly alleviated when ADI was used in the GT patients. However, other adverse events such as nephrotoxicity, diarrhea, alopecia, and neurotoxicity did not significantly differ between the two groups. Overall, these results suggest that the combination of conventional and ADI treatment is more effective than conventional treatment alone.     

Studies on the regioselectivity and kinetics of the action of trypsin on proinsulin and its derivatives using mass spectrometry

Human M-proinsulin was cleaved by trypsin at the R³¹R³²–E³³ and K⁶⁴R⁶⁵–G⁶⁶ bonds (B/C and C/A junctions), showing the same cleavage specificity as exhibited by prohormone convertases 1 and 2 respectively. Buffalo/bovine M-proinsulin was also cleaved by trypsin at the K⁵⁹R⁶⁰–G⁶¹ bond but at the B/C junction cleavage occurred at the R³¹R³²–E³³ as well as the R³¹–R³²E³³ bond. Thus, the human isoform in the native state, with a 31 residue connecting C-peptide, seems to have a unique structure around the B/C and C/A junctions and cleavage at these sites is predominantly governed by the structure of the proinsulin itself. In the case of both the proinsulin species the cleavage at the B/C junction was preferred (65%) over that at the C/A junction (35%) supporting the earlier suggestion of the presence of some form of secondary structure at the C/A junction. Proinsulin and its derivatives, as natural substrates for trypsin, were used and mass spectrometric analysis showed that the k_cat./K_m values for the cleavage were most favourable for the scission of the bonds at the two junctions (1.02 ± 0.08 × 10⁵ s^− 1 M^− 1) and the cleavage of the K²⁹–T³⁰ bond of M-insulin-RR (1.3 ± 0.07 × 10⁵ s^− 1 M^− 1). However, the K²⁹–T³⁰ bond in M-insulin, insulin as well as M-proinsulin was shielded from attack by trypsin (k_cat./K_m values around 1000 s^− 1 M^− 1). Hence, as the biosynthetic path follows the sequence; proinsulin → insulin-RR → insulin, the K²⁹–T³⁰ bond becomes shielded, exposed then shielded again respectively.     

Soil tillage reduces arthropod biodiversity and has lag effects within organic and conventional crop rotations

Crop rotation systems in organic and conventional farming systems differ in crop types, management and duration. However, changes in arthropod communities over the entire rotation system are poorly understood, as many studies have surveyed only single years or have not covered the entire rotation period. Here, we describe changes in arthropods in two contrasting systems at a split organic‐conventional farm: an 8‐year organically managed rotation with five crops and a 5‐year conventionally managed rotation with three crops. Arthropods were classified into three functional groups, representing epigeal predators, foliar predators/parasitoids and herbivores/pollinators. Epigeal predators were particularly reduced by soil tillage which occurred annually in the conventional rotation, but was intermittent in the organic. Arthropods were most abundant on the conventional rotation, but most taxonomically diverse on the organic. In the conventional system, all functional groups showed a cyclical change in their taxonomic composition that closely matched the crop rotation sequence, whereas in the organic rotation, the cycle was less clear. Whilst the current year's crop type was the major determinant of arthropod community composition, there was a significant “lag effect” for many taxa from the preceding year's crop. Our results suggest that both the amounts of soil tillage (e.g., in no‐till systems) and crop rotation order have major impacts on arthropods in agroecosystems. Rotations with excessive soil tillage are likely to reduce the abundance of some groups of beneficial arthropods, especially epigeal predators.     

不同水稻品系幼苗对砷（As）的耐性、吸收及转运

利用琼脂培养基模拟水稻生长的厌氧环境,研究了As对不同水稻品系幼苗生长的影响以及As在其体内的积累及转运特性.结果表明,不同浓度(0～4.0mg/L)的As对供试水稻品系根部干物质积累无显著影响(P>0.05).杂交稻与糯稻的地上部干物质积累随基质中As浓度的增加呈减小趋势,但低剂量的As(0.5mg/L)促进常规稻的生长.水稻地上部的As积累量随基质中As浓度的升高总体均呈增加趋势.水稻根系对As具有较强的吸收与累积能力.水稻根部As的积累量为156.31～504 03mg/kg,占总As含量的63.40～81.90%,远远高于其地上部As的积累量.相比于其它两个品系,糯稻的生物量积累高,耐性指数较大,根部及地上部对As的积累量较低,因此更适合种植在As污染土壤.     

Rapid Identification of Viable Bacterial Spores Using a Fluorescence Method

A method has been devised to differentiate viable and nonviable bacterial spores. “Germination-like” changes are initiated in spores with performic acid and lysozyme. The germinated spores are stained with aqueous acridine orange, a fluorescent dye. Nonviable spores fluoresce lemon-green and viable spores orange-red. It is proposed that with the use of a membrane filter resistant to performic acid and lysozyme, the method may be used for spore enumeration in foods in about 4 hr compared to conventional plating methods, which usually require up to 72 hr.     

Neuroprotective and anticonvulsant effects of organic and conventional purple grape juices on seizures in Wistar rats induced by pentylenetetrazole

Epilepsy is the most common neurological disorder worldwide. Studies have shown that recurrent seizures may increase the concentration of reactive oxygen species, which can lead to oxidative stress and neuronal damage. These seizures result in substantial deleterious effects on an individual's health. Organic and conventional grape juices are rich in polyphenols, compounds with important antioxidant activity. However, these juices could have differences in their polyphenol content. The aim of this study was to investigate the neuroprotective and anticonvulsant effects of organic and conventional grape juice treatments in Wistar rats against pentylenetetrazole (a convulsant drug)-induced damage. In addition, we evaluated potential behavioral changes in rats treated with the juices and the polyphenolic profile of those samples. Animals (n=16 in each group) received treatment with saline, organic or conventional grape juice for 17 days. On the eighteenth day, behavioral changes were evaluated by an open field test. Afterwards, half of the rats from each group received pentylenetetrazole and were observed for 30 min to evaluate possible seizure characteristics. The animals were subsequently killed by decapitation and their hippocampus, cerebellum and cerebral cortex tissues were isolated. The results of this study showed that neither organic nor conventional grape juice altered the behavior parameters, and no statistical differences were observed in the seizure characteristics of the groups. Nevertheless, both juice types were able to protect from lipid and protein oxidative damage, decrease nitric oxide content and increase enzymatic (superoxide dismutase and catalase) and non-enzymatic (sulfhydryl protein) antioxidant defenses in brain tissues following pentylenetetrazole-induced seizures. In general, organic juice showed superior results in each test, probably due to its higher polyphenol content relative to conventional juice. These results indicate that grape juices can provide further insight into natural neuroprotective compounds and may lead to the development of new therapeutic strategies for epileptic patients.     

Carbon accumulation at depth in Ferralsols under zero‐till subtropical agriculture

Conservation agriculture can provide a low‐cost competitive option to mitigate global warming with reduction or elimination of soil tillage and increase soil organic carbon (SOC). Most studies have evaluated the impact of zero till (ZT) only on surface soil layers (down to 30 cm), and few studies have been performed on the potential for C accumulation in deeper layers (0–100 cm) of tropical and subtropical soils. In order to determine whether the change from conventional tillage (CT) to ZT has induced a net gain in SOC, three long‐term experiments (15–26 years) on free‐draining Ferralsols in the subtropical region of South Brazil were sampled and the SOC stocks to 30 and 100 cm calculated on an equivalent soil mass basis. In rotations containing intercropped or cover‐crop legumes, there were significant accumulations of SOC in ZT soils varying from 5 to 8 Mg ha^?1 in comparison with CT management, equivalent to annual soil C accumulation rates of between 0.04 and 0.88 Mg ha^?1. However, the potential for soil C accumulation was considerably increased (varying from 0.48 to 1.53 Mg ha^?1 yr^?1) when considering the soil profile down to 100 cm depth. On average the estimate of soil C accumulation to 100 cm depth was 59% greater than that for soil C accumulated to 30 cm. These findings suggest that increasing sampling depth from 30 cm (as presently recommended by the IPCC) to 100 cm, may increase substantially the estimates of potential CO₂ mitigation induced by the change from CT to ZT on the free‐draining Ferralsols of the tropics and subtropics. It was evident that that legumes which contributed a net input of biologically fixed N played an important role in promoting soil C accumulation in these soils under ZT, perhaps due to a slow‐release of N from decaying surface residues/roots which favored maize root growth.     

茉莉酸对棉花单宁含量和抗虫相关酶活性的诱导效应

以植物生长调节物茉莉酸(Jasmonic acid,JA)为诱导子,以常规棉为研究对象,探讨了外源茉莉酸对棉花幼苗单宁和蛋白酶抑制素以及其它抗虫相关酶活性诱导的浓度依赖性和持久性,讨论了棉花抗虫相关物质的抗虫效果.结果表明,0.01、0.1和1.0 mmol/L茉莉酸都能在2周内诱导棉花单宁和胰蛋白酶抑制素(Proteinase inhibitors,PIs)含量增加,诱导多酚氧化酶(Polyphenol oxidase,PPO)、苯丙氨酸解氨酶(Phenylalanine ammonia-lyase,PAL)、过氧化物酶(Peroxidase,POD)和过氧化氢酶(Catalase,CAT)活性升高.对3种浓度茉莉酸的诱导效应进行分析表明,0.1 mmol/L茉莉酸对于诱导PIs、PPO、POD和CAT最有效,0.1和1.0 mmol/L茉莉酸对于诱导棉花单宁和苯丙氨酸解氨酶等效,二者的诱导效应均高于0.01 mmol/L.对茉莉酸诱导抗性的持久性进行分析表明,最佳诱导效应发生的时间各不相同:POD活性在JA处理后第1天最高,随后呈下降趋势,PIs和单宁含量分别在JA处理后第7天和第14天达最大值;JA处理后第1天和第7天的PPO活性无明显差异,但明显高于第14天;JA处理后第7天和第14天的PAL活性无明显差异,但明显高于第1天;JA处理后第1、7和14天棉花叶片的CAT活性均无明显差异.以上结果表明,茉莉酸可通过增加棉叶单宁和PIs含量、提高棉叶PAL、PPO、POD和CAT活性等增强棉花幼苗的抗虫性.