Effect of efficient pectinolytic bacterial isolates on retting and fibre quality of jute

Four bacterial strains with high balanced polygalacturonase (PG) and pectin lyase (PNL), and xylanase with minimal cellulase activity were used in jute retting. The 16S rDNA analysis revealed that the organisms were: Bacillus sp. L6 (GQ891097), Bacillus pumilus strain EK-17 (GQ891098), B. pumilus strain Geo-03-422 (GQ891103) and B. pumilus strain IK-MB12-518F (GQ891105). The microorganisms in different combinations of consortia showed synergistic effect resulting in increased PG (35.52-46.61 IU/g cell wet weight), PNL (39.79-72.12 U/ml), xylanase (0.705-0.840 μmol/ml/min) and little cellulase (0-0.153 μmol/ml/min) activities. The organisms in different combinations of consortia reduced the retting period from 11 to 13 days as compared to 19 days in the control. Microbial inoculation produced remarkable improvement in jute fibre strength (26.62-28.91 g/tex) and fineness (2.76-2.92 tex) over control. The pH of the post-retting water samples became acidic, and the electrical conductivity (Ec), chemical oxygen demand (COD) and hardness increased. The organisms could be adopted in industrial application for extraction of jute fibre.     

杧果炭疽病菌 3 个果胶裂解酶基因序列特征及受漆酶基因Lac1 影响的分析

果胶裂解酶是炭疽菌在侵染寄主过程中降解寄主细胞壁的一类重要水解酶。本研究在杧果炭疽病菌中克隆获得了 3 个果胶裂解酶基因 Cgpel1、Cgpel2 和 Cgpel3,DNA 全长分别为 1 037、1 498、1 089 bp,cDNA 全长分别为 975、 1 380、978 bp,分别编码 324、459、325 个氨基酸,均含 1 个果胶裂解酶保守结构域,均有典型的信号肽,不存在跨膜结构。其二级结构中 α-螺旋分别占 16.05%、20.26%、16.92%,延伸链分别占 28.09%、21.79%、29.54%,β-转角分别占 5.86%、8.93%、7.38%,无规则卷曲分别占 50.00%、49.02%、46.15%。Cgpel1、Cgpel2 和 Cgpel3 的氨基酸序列分别与 C. tofieldiae（KZL77240.1）、草莓炭疽菌（C. gloeosporioides）（XP-007274932.1）、C. incanum（KZL84476.1）果胶裂解酶序列相似度在 93%以上。qRT-PCR 分析发现,3 个果胶裂解酶基因在整个侵染过程中均持续高效表达,但在漆酶基因 Lac1 敲除突变体中,表达均下降约 90%。可见 Cgpel1、Cgpel2 和 Cgpel3 是果胶裂解酶基因家族成员,序列差异较大,在侵染过程中起着重要的作用,且其表达受漆酶基因 Lac1 影响。     

Purification and Characterization of a Novel Alginate Lyase from a Marine Streptomyces Species Isolated from Seaweed

Alginate, a natural polysaccharide derived from brown seaweed, is finding multiple applications in biomedicine via its transformation through chemical, physical, and, increasingly, enzymatic processes. In this study a novel alginate lyase, AlyDS44, was purified and characterized from a marine actinobacterium, Streptomyces luridiscabiei, which was isolated from decomposing seaweed. The purified enzyme had a specific activity of 108.6 U/mg, with a molecular weight of 28.6 kDa, and was composed of 260 amino acid residues. AlyDS44 is a bifunctional alginate lyase, active on both polyguluronate and polymannuronate, though it preferentially degrades polyguluronate. The optimal pH of this enzyme is 8.5 and the optimal temperature is 45 °C. It is a salt-tolerant alginate lyase with an optimal activity at 0.6 M NaCl. Metal ions Mn²⁺, Co²⁺, and Fe²⁺ increased the alginate degrading activity, but it was inhibited in the presence of Zn²⁺ and Cu²⁺. The highly conserved regions of its amino acid sequences indicated that AlyDS44 belongs to the polysaccharide lyase family 7. The main breakdown products of the enzyme on alginate were disaccharides, trisaccharides, and tetrasaccharides, which demonstrated that this enzyme acted as an endo-type alginate lyase. AlyDS44 is a novel enzyme, with the potential for efficient production of alginate oligosaccharides with low degrees of polymerization.     

烟草野火病菌(Pseudomonas syringae pv. tabaci)对烟草细胞内5种防御酶系统的影响

定期测定了感病品种红花大金元接种烟草野火病菌后叶片内5种酶活性的动态变化，研究结果表明：烟草接种病菌后，SOD活性先上升，后在8d下降，低于对照；POD活性接种后在1d略低于对照，后上升较快，10d达到高峰，此后一直高于对照；PPO活性在接种后1d低于对照15.8%，但此后上升，16d达到高峰，18d下降低于对照；CAT活性变化与POD相似，接种1d低于对照，但此后一直高于对照，并于6d达到高峰，10d虽有所下降，但接着升高；PAL活性与CAT、POD变化相似，接种后1d活性低于对照28.3%，其后上升，10d达到高峰，是对照的2.11倍，并维持较高的水平。     

中生菌素对水稻白叶枯病的防治机制

对不同白叶枯病抗性水稻品种用200μg/ml中生菌素55℃温汤药液浸种，自然降温，秧田3—4叶期和移栽前5d各用30μg/ml中生菌素处理后，于成株期剪叶接种白叶枯病菌。结果表明中生菌素前期处理，于成株期接种白叶枯病菌时，高抗、中等抗性和感病品种中过氧化物酶、苯丙氨酸解氨酶和多酚氧化酶3种酶活性都有不同程度的提高，且以中等抗性品种酶活提高最多，接种24和48h，3种酶活性较清水对照分别增加26．92％、26．74％、24．06％和7．09％、1．31％、1．60％。盆栽试验表明，中生菌素对中抗品种的防治效果最好，达58．4％。说明中生菌素对水稻防御酶活性的诱激作用是其防治白叶枯病的机制之一。     

Influence of Abscisic Acid and the Abscisic Acid Biosynthesis Inhibitor, Norflurazon, on Interactions Between Phytophthora sojae and Soybean (Glycine max)

A comparison was made of the effects of abscisic acid (ABA) and the ABA biosynthesis inhibitor, norflurazon on the interaction between soybean leaves and Phytophthora sojae. Inoculation of leaves of cv. Harosoy resulted in a compatible interaction typified by the presence of spreading, water soaked lesions with ill-defined margins while inoculation of cv. Haro 1272 resulted in an incompatible interaction with lesions restricted to the inoculation site. Activity of phenylalanine ammonia lyase (PAL) slowly increased in the compatible interaction but in the incompatible interaction there was a rapid rise in activity within 8h after inoculation. When Haro 1272 plants were treated with ABA the normally incompatible interaction with race 1 was changed to what resembled a compatible interaction and activity of PAL was reduced to control levels. There was no visible effect on the compatible combination. In contrast when plants of cv. Harosoy were treated with norflurazon the normally compatible interaction with race 1 was changed to that which resembled an incompatible interaction and PAL activity increased to high levels rapidly. There was no effect of norflurazon on the incompatible interaction of cv. Haro 1272 with race 1. Stomata on leaves of cv. Harosoy treated with norflurazon closed within 2h of inoculation resembling the response of stomata in normal incompatible interactions but not compatible interactions where stomata remained open. On leaves of cv. Harosoy treated with norflurazon at sites 3 and 20mm from the inoculation point stomata also closed. These results extend and confirm the idea that ABA is a molecule that may regulate the outcome of the interaction between soybeans and P. sojae.     

植物激活蛋白对玉米抗病相关酶活性的影响

以稀释1000倍的植物激活蛋白粗提液处理玉米,以稀释1000倍的pH6.8磷酸缓冲液作对照,分别在处理后1、4、7、10、13和16d测定了玉米中苯丙氨酸解氨酶、过氧化物酶、几丁质酶和多酚氧化酶活性。结果表明:在不同时期处理后4种酶活性比对照有所提高,说明植物激活蛋白能诱导玉米抗病性。     

NMR Analyses of the Enzymatic Degradation End-Products of Diabolican: The Secreted EPS of Vibrio diabolicus CNCM I-1629

Diabolican, or HE800, is an exopolysaccharide secreted by the non-pathogenic Gram-negative marine bacterium Vibrio diabolicus (CNCM I-1629). This polysaccharide was enzymatically degraded by the Bacteroides cellulosilyticus WH2 hyaluronan lyase. The end products were purified by size-exclusion chromatography and their structures were analyzed in depth by nuclear magnetic resonance (NMR). The oligosaccharide structures confirmed the possible site of cleavage of the enzyme showing plasticity in the substrate recognitions. The production of glycosaminoglycan-mimetic oligosaccharides of defined molecular weight and structure opens new perspectives in the valorization of the marine polysaccharide diabolican.     

Characterization of a Novel Alginate Lyase with Two Alginate Lyase Domains from the Marine Bacterium Vibrio sp. C42

Alginate is abundant in the cell walls of brown algae. Alginate lyases can degrade alginate, and thus play an important role in the marine carbon cycle and industrial production. Currently, most reported alginate lyases contain only one functional alginate lyase domain. AlyC8 is a putative alginate lyase with two alginate lyase domains (CD1 and CD2) from the marine alginate-degrading strain Vibrio sp. C42. To characterize AlyC8 and its two catalytic domains, AlyC8 and its two catalytic domain-deleted mutants, AlyC8-CD1 and AlyC8-CD2, were expressed in Escherichia coli. All three proteins have noticeable activity toward sodium alginate and exhibit optimal activities at pH 8.0–9.0 and at 30–40 °C, demonstrating that both CD1 and CD2 are functional. However, CD1 and CD2 showed opposite substrate specificity. The differences in substrate specificity and degradation products of alginate between the mutants and AlyC8 demonstrate that CD1 and CD2 can act synergistically to enable AlyC8 to degrade various alginate substrates into smaller oligomeric products. Moreover, kinetic analysis indicated that AlyC8-CD1 plays a major role in the degradation of alginate by AlyC8. These results demonstrate that AlyC8 is a novel alginate lyase with two functional catalytic domains that are synergistic in alginate degradation, which is helpful for a better understanding of alginate lyases and alginate degradation.