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纤维素酶催化水解和氧化机制的研究进展   总被引:8,自引:0,他引:8  
人类对纤维素的开发与利用还非常有限,未能完全搞清天然纤维素的生物降解机制是其一个重要原因.自从C1-Cx假说提出以来,又有较多的降解理论出现对其改进,纤维素酶水解机制不能完全解释天然纤维素的降解,大量文献证实天然纤维素降解存在氧化机制.本文主要介绍水解与氧化机制中具代表性酶协同降解模型、纤维二糖水解酶(CBH)协同作用假说、羟基自由基的氧化机制和纤维二糖脱氢酶途径等不同降解理论,将水解和氧化两种机制结合起来给出其降解途径,也许能够较为容易理解纤维素的降解具体过程.  相似文献   
2.
刘沙沙  李静梅  石波  梁平  李超 《中国农业科学》2013,46(11):2345-2352
【目的】探索商品化β-葡聚糖酶水解小麦秸秆制备纤维寡糖的最佳条件,为纤维寡糖的工业化生产提供科学依据。【方法】采用浓硫酸和浓盐酸混酸降解微晶纤维素制备纤维寡糖混合物,通过分离纯化得到聚合度为2—5的纤维寡糖单一组分。用所得纤维二糖作为标准品测定小麦秸秆酶解产物中的纤维二糖,并以纤维二糖含量和小麦秸秆转化率为衡量指标,研究酶解反应温度、pH、反应酶底比E/S、反应底物浓度、反应时间对寡糖生成的影响。【结果】采用活性炭柱层析法得到了高纯度的聚合度2-5的纤维寡糖单一组分;当酶解反应温度为50℃、pH为5.5、E/S为0.4、底物浓度为2%、时间10 h时酶解效果最好,小麦秸秆总转化率达到55.57%,纤维二糖得率为148.15 mg•g-1秸秆。【结论】活性炭柱层析可以很好地分离纤维寡糖,得到了聚合度为2—5的纤维寡糖单一组分;酶解小麦秸秆总转化率及纤维二糖得率均提高。  相似文献   
3.
为探究水杨酸、壳聚糖、麦角甾醇和纤维二糖4种化合物诱导马铃薯抗晚疫病的效果及其作用机理,采用整株喷雾法、挑战接种法、菌丝生长速率法分析4种化合物对马铃薯晚疫病的预防作用及对其病原菌致病疫霉Phytophthora infestans的离体抑制效果,利用实时荧光定量PCR技术测定4种化合物诱导后马铃薯病程相关蛋白和防御酶基因的调控表达情况。结果显示,水杨酸、壳聚糖、纤维二糖和麦角甾醇分别处理马铃薯植株24 h后接种致病疫霉,在多个浓度下对晚疫病均有不同程度的诱导抗性,其中在1 000 μmol/L浓度(壳聚糖浓度为1 000 mg/L)时诱导抗性最好,较对照显著提高,诱抗效果分别为54.73%、61.99%、59.73%和48.59%。壳聚糖对马铃薯晚疫病的诱抗效果高于其它3种化合物,且只有壳聚糖能显著抑制致病疫霉的菌丝生长和孢子囊形成,说明壳聚糖除了能诱导作物产生抗病性外,还具有直接抑菌作用。4种化合物诱导后马铃薯病程相关蛋白基因PR1、过氧化物酶基因POD和多酚氧化酶基因PPO的表达量在施用早期均显著升高。表明这些化合物能诱导马铃薯对晚疫病产生抗性,可能与马铃薯体内过氧化物酶、多酚氧化酶的活性和抗病信号传导途径关键基因PR1的表达有关。  相似文献   
4.
Tests of several fumigants which control Phytophthora cinnamomi Rands under field and glasshouse conditions are reported. The soil fumigant basamid is the cheapest effective treatment available, at present. The implications of failure to control Phytophthora cinnamomi for forestry, agriculture and the indigenous flora are briefly discussed.  相似文献   
5.
A semi-selective medium for isolation of Xanthomonas axonopodis pv. vignicola from cowpea (Vigna unguiculata) plant and soil samples was developed. Twelve carbon and five nitrogen sources were tested with four strains of X. axonopodispv.vignicola, and 25 antibiotics were screened against saprophytes. -cellobiose (10g) was selected as the optimal carbon source. Among the antibiotics, cefazoline inhibited growth of most of the saprophytes with little effect on strains of the pathogen. ,-methionine enhanced growth of X. axonopodispv.vignicola. Boric acid along with ammonium chloride suppressed growth of Pseudomonas fluorescens. The semi-selective medium designated as cefazoline-cellobiose-methionine (CCM) medium contained K2HPO4 1.34g, KH2PO4 0.4g, MgSO4 0.3g, H3BO3 0.2g, NH4Cl 1.0g, -cellobiose 10g, cycloheximide 0.2g, ,-methionine 1.0g, cefazoline 10mg and agar 14g per l of water (pH 7.2). Colonies of X. axonopodispv.vignicola on CCM medium were whitish, round, raised and 0.2–1.8mm in diameter 96h after incubation. CCM medium generally inhibited growth of Pantoea agglomerans, Bacillus subtilis and saprophytes isolated from cowpea leaves. Colonies of Pseudomonas fluorescens and a saprophytic bacterium, which were not completely suppressed by CCM, could be differentiated from X. axonopodispv.vignicola by their smaller size and different color. The CCM medium proved useful for isolation of X. axonopodispv.vignicola from cowpea plant and soil samples. This is the first report of a semi-selective medium developed for detection of X. axonopodispv.vignicola.  相似文献   
6.
This experiment was designed to investigate the effects of different concentrations of cellobiose (CB) or a twin strain of Saccharomyces cerevisiae live cells (YST) (20, 40 and 60 mg/60 mL), and CB + YST (60 + 20, 60 + 40, 60 + 60 mg/60 mL) on mixed ruminal microorganism fermentation in vitro. Ruminal fluid was collected from a cow, mixed with phosphate buffer (1:2) and incubated (60 mL) anaerobically at 38°C for 24 h with or without supplement plus 400 mg (dry matter [DM] basis) substrate (hay plus concentrate, 1.5:1). The medium pH numerically decreased with CB and CB + YST, but was unchanged with YST. The total volatile fatty acid and proportion of propionate increased (P < 0.05) in all cases. The proportion of acetate decreased (P < 0.05) with CB and CB + YST, but increased (P < 0.05) with YST and that of butyrate increased (P < 0.05) with CB and CB + YST, but decreased (P < 0.05) with YST. Ammonia‐N decreased (P < 0.05) with CB and CB + YST, but was unchanged with YST. The number of protozoa was unchanged, and that of cellulolytic bacteria increased (P < 0.05) in all cases. Total gas production increased (P < 0.05) in all cases. Methane decreased, hydrogen was unchanged by YST and both gases were unchanged by CB and CB + YST. The in vitro disappearance of DM and neutral detergent fiber increased (P < 0.05) by 11.2% and 8.9%, 9% and 8.5%, and 12.1% and 10.2% in the case of CB, YST and CB + YST, respectively. Therefore, the dietary supplementation of CB and/or YST may improve ruminal fermentation and digestibility.  相似文献   
7.
牛瘤胃产纤维二糖磷酸化酶细菌的筛选及产酶条件研究   总被引:1,自引:0,他引:1  
为了获得产新型纤维二糖磷酸化酶(CBP)的菌株,利用选择性培养基从牛瘤胃液中逐步分离纯化,筛选能以纤维二糖为唯一碳源生长的菌株,分别检测所筛选菌株的β-葡萄糖苷酶(βG)和CBP活力,并对其纤维二糖的利用方式(水解途径或磷解途径)进行鉴定,获得产CBP的菌株,命名为BY-a。采用形态学观察、生理生化试验,结合16S r DNA序列分析,鉴定菌株BY-a为克雷伯氏菌属(Klebsiella)。对菌种产CBP的温度、时间、最适装液量和溶氧量进行初步优化,结果显示,BY-a菌株在装液量为30mL(250mL)、37℃振荡培养24 h时,产CBP活力最高。  相似文献   
8.
以蒸汽爆破玉米秸秆酶水解液为底物,选取温度、pH和酶用量为自变量,水解液中的葡萄糖含量为响应值,利用Box-Benhnken中心组合试验和响应面(RSM)分析法对β-葡萄糖苷酶均相水解条件进行优化。结果表明:水解温度和酶用量对葡萄糖含量影响显著;β-葡萄糖苷酶均相水解纤维二糖的最佳工艺为:温度52℃,pH4.0,酶用量1 IU/mL。在此条件下,3次验证实验的平均葡萄糖浓度为52.81 mg/mL。  相似文献   
9.
A genomic library of Xanthomonas oryzae pv. oryzae (X. o. pv. oryzae) T7174 was screened for 4-methylumbelliferyl β-D-glucoside-hydrolyzing (MUGase) activity. In subcloning of one of the MUGase-positive clones, an approximately 4.2-kb SacI-SphI fragment conferred not only MUGase activity but also 4-methylumbelliferyl β-D-cellobioside-hydrolyzing (MUCase) activity. Sequence analysis showed that the fragment contained an ORF of 2951 bp. The conceptual ORF product was significantly homologous with 1,4-β-D-glucan glucohydrolase D (CELD) from Pseudomonas fluorescens subsp. cellulosa, and was named CELDXo. Cell fractionation experiments suggested that CELDXo is localized in the cell-envelope fraction. We constructed a CELDXo-deficient mutant (74ΔCELD) from X. o. pv. oryzae. Little MUCase activity was detected in the cell-envelope fraction prepared from the mutant. The mutant 74ΔCELD did not grow in synthetic medium containing cellobiose as the sole sugar source. On the other hand, growth in rice leaves and pathogenicity of the mutant and the parental strain did not differ. These results suggested that CELDXo is involved in cellobiose utilization of X. o. pv. oryzae but that the gene is not required for bacterial growth in rice leaves. Received 16 February 2001/ Accepted in revised form 11 April 2001  相似文献   
10.
In this in vitro study, the modified Hohenheim gas test (HGT) was applied to determine fermentation activity and bacterial composition of pig's faecal microbial inoculum using different concentrations of cellobiose. Incubation procedures included normal buffered and osmotic stress conditions (elevated medium salinity). After 24 hr of fermentation, production of gas, ammonia and short‐chain fatty acid (SCFA) was measured, and the gene copy numbers of total bacteria, Lactobacillus spp., Bifidobacterium spp., Roseburia spp., Clostridium Cluster IV spp. and Enterobacteriaceae were analysed using real‐time polymerase chain reaction. There was a significant reduction in gas production after 24 hr when comparing osmotic stress conditions with normal buffered conditions. Under osmotic stress, increasing cellobiose concentrations linearly increased gas production (< .001), while ammonia, acetic acid and isobutyric acid concentrations decreased (< .001, = .012, = .035 respectively). Under normal buffered conditions, Roseburia spp. gene copies linearly increased with increasing cellobiose concentrations (= .048). Lactobacillus spp. and Bifidobacterium spp. numbers were higher under osmotic stress (< .001) compared to normal conditions. Results might point towards a positive impact of cellobiose supplementation on gut health especially under osmotic stress conditions.  相似文献   
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