Characterization of Full-Biodegradable Starch-PVA-Polyester Film

IntroductionConventional polymers such as polyethyleneand polypropylene persistfor many years after landdisposal.Furthermore,plastics are often soiled byfood and other biological substances,making phys-ical recycling of those materials impractical andgenerally undesirable. In contrast,biodegradablepolymers disposed in bioactive environment are de-graded by the enzymatic action of microorganismssuch as bacteria,fungi,and algae.The worldwideconsumption of biodegradable polymers increasedfrom1.4×107kg in ...

Comparison of two marine sponge-associated Penicillium strains DQ25 and SC10: differences in secondary metabolites and their bioactivities

Two strains of Penicillium, DQ25 and SC10, isolated from marine sponge Haliclona angulata (Bowerbank) and Hymeniacidon sp. respectively, were subjected to stationary cultivation under GYP medium for 30 days. The fermentation extracts were undergone bioactivities assays against human pathogens, phytopathogenic fungi and brine shrimp (Artemia salina). Bioassays-guided compounds isolation was performed by Silica gel columns and Sephadex LH-20 chromatography. Spectroscopic methods were used to structures elucidation of the compounds. Results showed the activities of secondary metabolites of strain DQ25 were generally stronger than that of strain SC10. Major bioactive molecules isolated from strain DQ25 were a 1,4-naphthoquinone derivative and an unidentified alkaloid. The two components were not isolated from the extract of strain SC10. ITS sequences revealed that these two species have the greatest similarity with Penicillium vinaceum and Penicillium granulatum respectively.

High level expression, purification, and characterization of the shrimp antimicrobial peptide, Ch-penaeidin, in Pichia pastoris

Penaeidins, members of a new family of antimicrobial peptides constitutively produced and stored in the haemocytes of penaeid shrimp, display antimicrobial activity against bacteria, and fungi. Here, a DNA sequence encoding the mature Ch-penaeidin peptide was cloned into the pPIC9K vector and transformed into Pichia pastoris. The transformed cells were screened for multi-copy plasmids using increasing concentrations of G418. Positive colonies carrying chromosomal integrations of the Chp gene were identified by phenotype and PCR. When transformed cells were induced with methanol, SDS-PAGE and Western blotting revealed the production of a similar to6100 Da recombinant CHP (rCHP) expression product. Large scale expression revealed that rCHP was produced at 108 mg/L under optimal conditions in the highest Chp-producing P. pastoris clone. The antimicrobial activities of rCHP were studied by liquid phase analysis, which revealed that rCHP exhibited activities against some Gram-negative and Gram-positive bacteria, but had a relatively low activity against some fungi. Purification of rCHP by cation exchange chromatography and subsequent automated amino acid sequencing revealed the presence of four additional amino acids (YVEF) at the N-terminus that belonged to the cleaved fusion signal peptide; these residues may account for the observed decrease in antifungal activity. Together, these observations indicate that rCHP is an effective antimicrobial peptide that can be successfully produced at high levels in the yeast, and therefore may be a potential antimicrobial candidate for practical use. (C) 2004 Elsevier Inc. All rights reserved.

Ergosterimide, a new natural Diels-Alder adduct of a steroid and maleimide in the fungus Aspergillus niger

Ergosterimide (1), a natural Diels-Alder adduct of ergosteroid and maleimide, was characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from the marine brown alga Colpomenia sinuosa. In addition, four known steroids including (22E,24R)-ergosta-5,7,22-trien-3 beta-ol (2), (22E,24R)-ergosta-4,6,8(14),22-tetraen-3one (3), (22E,24R)-5 alpha,8 alpha-epidioxyergosta-6,22-dien-3 beta-ol (4), and (22E,24R)-ergosta-7,22dien-3 beta,5 alpha,6 beta-triol. (5) were also isolated and identified. The structures of these compounds were elucidated by extensive analysis of 1D and 2D NMR and IR spectra and MS data. The plausible biosynthetic pathway of 1 was also discussed. To the best of our knowledge, 1 is the first natural Diels-Alder adduct of steroid and maleimide reported so far. (C) 2007 Elsevier Inc. All rights reserved.

CfLGBP, a pattern recognition receptor in Chlamys farreri involved in the immune response against various bacteria

Lipopolysaccharide and beta-1, 3-glucan binding protein (LGBP) is a kind of pattern recognition receptor, which can recognize and bind LPS and beta-1, 3-glucan, and plays curial roles in the innate immune defense against Gram-negative bacteria and fungi. In this study, the functions of LGBP from Zhikong scallop Chlamys farreri performed in innate immunity were analyzed. Firstly, the mRNA expression of CfLGBP in hemocytes toward three typical PAMPS stimulation was examined by realtime PCR. It was up-regulated extremely (P < 0.01) post stimulation of LPS and beta-glucan, and also exhibited a moderate up-regulation (P < 0.01) after PGN injection. Further PAMPs binding assay with the polyclonal antibody specific for CfLGBP proved that the recombinant CfLGBP (designated as rCfLGBP) could bind not only LPS and beta-glucan, but also PGN in vitro. More importantly, rCfLGBP exhibited obvious agglutination activity towards Gram-negative bacteria Escherichia coil, Gram-positive bacteria Bacillus subtilis and fungi Pichia pastoris. Taking the results of immunofluorescence assay into account, which displayed CfLGBP was expressed specifically in the immune cells (hemocytes) and vulnerable organ (gill and mantle), we believed that LGBP in C farreri, serving as a multi-functional PRR, not only involved in the immune response against Gram-negative and fungi as LGBP in other invertebrates, but also played significant role in the event of anti-Gram-positive bacteria infection. As the first functional research of LGBP in mollusks, our study provided new implication into the innate immune defense mechanisms of C. farreri and mollusks. (C) 2010 Elsevier Ltd. All rights reserved.

红树林植物木榄和一株海藻内生真菌的化学成分研究

本论文对民间药用红树植物木榄（Bruguiera gymnorrhiza）中的次生代谢产物进行了研究，同时还对分离自似瘤凹顶藻（Laurencia similis Namet Saito）的内生真菌EN21（Exophiala oligosperma）进行了化学成分研究。对分离得到的部分化合物进行了初步的DPPH自由基清除活性和抗菌活性筛选。 采用常规的硅胶柱层析、反相硅胶柱层析、制备薄层层析、凝胶Sephadex LH-20 柱层析、半制备型高效液相色谱（HPLC）以及重结晶等手段，从木榄（Bruguiera gymnorrhiza）中分离得到23个化合物；从EN21（Exophiala oligosperma）中分离得到23个化合物。利用一维、二维核磁、质谱、元素分析等各种现代波谱技术及化学反应方法，确定了木榄中20个化合物的结构，包括黄酮单体化合物5个，三萜类化合物11个，甾体化合物2个，长链烷烃2个，其中4个化合物为首次从木榄中报道的黄酮；确定了EN-21中20个化合物的结构，其中脑苷脂1个，甾体化合物7个，芳环化合物3个，三萜类化合物2个，包括1个新天然产物。本文系首次报道Exophiala oligosperma 的化学成分。 对从木榄中分离得到的部分化合物进行了DPPH自由基清除活性筛选， 发现黄酮类化合物BRG-1、BRG-2、BRG-3 、BRG-4显示了较强的DPPH自由基清除活性，而三萜类化合物则未表现出DPPH自由基清除活性。抗菌筛选实验中所有测试样品未显示出活性。 研究结果充实了红树林植物和海藻内生真菌代谢产物的内容，为其有效利用提供了一定的科学依据。

黄槿内生真菌赤散囊菌及红海榄的次生代谢产物及其生物活性研究

由于生长环境的特殊性，红树林及其内生真菌的代谢产物在化学类型和生物活性方面都具有多样性，因此对其代谢产物的研究引起了人们越来越多的关注。本论文以菌丝体生物量、代谢产物量等指标及薄层色谱分析、高效液相色谱分析、抗菌活性测试等筛选手段对来源于我国海南红树植物的九株内生真菌在四种不同液体培养基上的静置发酵产物进行了综合评价，并从中选择了来源于半红树植物黄槿（Hibiscus tiliaceus）的内生真菌G2——赤散囊菌（Eurotium rubrum）进行了30 L规模发酵（采用PDB培养基）和次生代谢产物的研究，对分离得到的部分化合物进行了初步的生物活性评价。此外，本论文还对海南真红树植物红海榄（Rhizophora stylosa Griff.）的化学成分进行了研究，并对分离得到的部分化合物进行了二苯代苦味酰自由基（DPPH）清除活性的评价。 采用常规的硅胶柱层析、Sephadex LH-20柱层析、反相硅胶柱层析、制备薄层层析（pTLC）、重结晶等分离手段，分离纯化得到单体化合物。综合运用现代波谱技术 (IR、UV、MS、1D-NMR 和 2D-NMR) 以及与标准品或文献比对鉴定单体化合物的结构。从G2菌丝体和发酵液的合并提取物中鉴定了45个化合物的结构，其中13个为新化合物，结构类型包括6个苯甲醛类化合物（ER1*~ER6*）、4个蒽醌类化合物（ER15*~ ER18*）和3个含吲哚的二酮哌嗪生物碱类化合物（ER27*~ER29*）。 对以上分离鉴定的部分单体化合物进行了DPPH自由基清除活性、拒食杀虫活性、抗细菌活性以及体外细胞毒活性的初步评价。新化合物ER15*和三个已知化合物ER20、ER39和ER40都表现很强的DPPH自由基清除活性。化合物ER15*还表现较好的拒食杀虫活性，而化合物ER5*和ER18*不但没有杀虫活性，反而能促进幼虫的生长。所测试的化合物只有ER15*和ER18*表现出微弱的抗金黄色葡萄球菌活性。所测试的化合物对A-549、HL-60和P-388细胞株均未表现出有意义的体外细胞毒活性。 从红海榄枝条的提取物中分离鉴定了29个化合物，其中2个为新化合物，包括1个三萜酯（RS1*）和1个黄烷醇类化合物（RS12*）；另有1个三萜酯（RS5）和1个黄烷醇类化合物（RS11）作为新的天然产物被分离鉴定。 对从红海榄提取物乙酸乙酯相和正丁醇相分离得到的部分单体化合物进行了DPPH自由基清除活性的研究。黄烷醇类化合物RS16和RS17显示最强的活性。另外，实验结果说明黄烷醇类化合物的DPPH自由基清除活性与其分子中所含的羟基数目有一定关系，而且若芳香环上有多个邻位酚羟基，则该化合物的活性将增强。本论文实验结果为海南红海榄植物资源的利用提供了化学成分及抗氧化活性方面的科学依据。

Antimicrobial activity of hydroxylbenzenesulfonailides derivatives of chitosan, chitosan sulfates and carboxymethyl chitosan

Chitosan, carboxymethyl chitosan (CIVICS) and chitosan sulfates (CSS) with different molecular weight were modified by reacting with 4-hydroxyl-5-chloride-1,3-benzene-disulfo-chloride or 2-hydroxyl-5-chloride-1,3 -benzene-disulfo-chloride to give 12 kinds of new hydroxylbenzenesulfonailides derivatives of them. The preparation conditions of the derivatives were discussed in this paper, and their structures were characterized by FT-IR and C-13 NMR spectroscopy. The solubility of the derivatives was measured in the experiment. In addition, their antimicrobial activities against four bacteria and five crop-threatening pathogenic fungi were tested in the experiment. Besides, the rule and mechanism of their antibacterial activities were discussed in this paper. (C) 2009 Published by Elsevier B.V.

Preparation, characterization and antifungal properties of 2-(alpha-arylamino phosphonate)-chitosan

In this paper, 20 kinds of different 2-(alpha-arylamino phosphonate)-chitosan (2-alpha-AAPCS) were prepared by different Schiff bases of chitosan (CS) reacted with di-alkyl phosphite in benzene solution. The structures of the derivatives (2-alpha-AAPCS) were characterized by FT-IR spectroscopy and elemental analysis. In addition, the antifungal activities of the derivatives against four kinds of fungi were evaluated in the experiment. The results indicated that all the prepared 2-alpha-AAPCS had a significant inhibiting effect on the investigated fungi when the derivatives concentration ranged from 50 to 500 mu g mL(-1). Furthermore, the antifungal activities of the derivatives increased with increasing the molecular weight and concentration. And the antifungal activities of the derivatives were affected by their dimensional effect and charge density. Besides, the rule and mechanism of the antifungal activities of them were discussed in this paper. (C) 2009 Elsevier B.V. All rights reserved.

Synthesis and antifungal properties of sulfanilamide derivatives of chitosan

Sulfanilamide derivatives of chitosan (2-(4-acetamido-2-sulfanimide)-chitosan (HSACS, LSACS), 2-(4-acetamido-2-sulfanimide)-6-sulfo-chitosan (HSACSS, LSACSS) and 2-(4-acetamido-2-sulfanimide)-6-carboxymethyl-chitosan (HSACMCS, LSACMCS)) were prepared using different molecular weights of chitosan (CS), carboxymethyl chitosan (CMCS) and chitosan sulfates (CSS) reacted with 4-acetamidobenzene sulfonyl chloride in dimethylsulfoxide solution. The structures of the derivatives were characterized by FT-IR spectroscopy and elemental analysis, which showed that the substitution degree of sulfanilamide group of HSACS, HSACSS, HSACMCS, LSACS, LSACSS and LSACMCS were 0.623, 0.492, 0.515, 0.576, 0.463 and 0.477, respectively. The solubility of the derivatives (pH < 7.5) was higher than that of chitosan (pH < 6.5). The antifungal activities of the derivatives against Aiternaria solani and Phomopsis asparagi were evaluated based on the method of Jasso et al. in the experiment. The results indicated that all the prepared sulfanilamide derivatives had a significant inhibiting effect on the investigated fungi in the polymer concentration range from 50 to 500 mu g mL(-1). The antifungal activities of the derivatives increased with increasing the molecular weight, concentration or the substitution degree. The sulfanilamide derivatives of CS, CMCS and CSS show stronger antifungal activities than CS, CMCS and CSS. (C) 2007 Elsevier Ltd. All rights reserved.

Synthesis of acyl thiourea derivatives of chitosan and their antimicrobial activities in vitro

Three different acyl thiourea derivatives of chitosan (CS) were synthesized and their structures were characterized by FT-IR spectroscopy and elemental analysis. The antimicrobial behaviors of CS and its derivatives against four species of bacteria (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Sarcina) and four crop-threatening pathogenic fungi (Alternaria solani, Fusarium oxysporum f. sp. vasinfectum, Colletotrichum gloeosporioides (Penz.) Saec, and Phyllisticta zingiberi) were investigated. The results indicated that the antimicrobial activities of the acyl thiourea derivatives are much better than that of the parent CS. The minimum value of MIC and MBC of the derivatives against E coli was 15.62 and 62.49 mu g/mL, respectively. All of the acyl thiourea derivatives had a significant inhibitory effect on the fungi in concentrations of 50 - 500 mu g/mL; the maximum inhibitory index was 66.67%. The antifungal activities of the chloracetyl thiourea derivatives of CS are noticeably higher than the acetyl and benzoyl thiourea derivatives. The degree of grafting of the acyl thiourea group in the derivatives was related to antifungal activity; higher substitution resulted in stronger antifungal activity. (c) 2007 Elsevier Ltd. All rights reserved.

Oxalate decarboxylase from Agrobacterium tumefaciens C58 is translocated by a twin arginine translocation system

Oxalate decarboxylases (OXDCs) (E.C. 4.1.1.2) are enzymes catalyzing the conversion of oxalate to formate and CO2. The OXDCs found in fungi and bacteria belong to a functionally diverse protein superfamily known as the cupins. Fungi-originated OXDCs are secretory enzymes. However, most bacterial OXDCs are localized in the cytosol, and may be involved in energy metabolism. In Agrobacterium tumefaciens C58, a locus for a putative oxalate decarboxylase is present. In the study reported here, an enzyme was overexpressed in Escherichia coli and showed oxalate decarboxylase activity. Computational analysis revealed the A. tumefaciens C58 OXDC contains a signal peptide mediating translocation of the enzyme into the periplasm that was supported by expression of signal-peptideless and full-length versions of the enzyme in A. tumefaciens C58. Further site-directed mutagenesis experiment demonstrated that the A. tumefaciens C58 OXDC is most likely translocated by a twin-arginine translocation (TAT) system.

中国黄土高原洛川剖面 630 kyr 以来的孢粉记录

It is known that global climate changed from the early Tertiary “Green House” to the Quaternary “Ice House” of cyclic glacial-interglacial climatic changes. Since the middle Pleistocene, the climate cycles changed from 40 kyr to 100 kyr, and the amplitudes of climatic fluctuations increased significantly. Therefore, it is important to study the climate changes since the middle Pleistocene. The loess-paleosol sequence in China is considered as one of the most continuous continental records of the last 2.58 Ma. Paleoclimatic and environmental changes have been widely extracted through various climatic parameters. However, the history of paleovegetation on the Loess Plateau still remains unclear. Did an extensive broadleaf forest ever exist on the Loess Plateau? Pollen preserved in the loess and paleosol provides a direct record for vegetation and paleoenvironmental change on the Plateau. However, because it is difficult to extract sufficient pollen grains from loess, the pollen record since the middle Pleistocene especially in the central part of the Chinese Loess Plateau has not been well studied. So we preliminarily focus on the palynological records of the loess-paleosol sequence spanning the last 630 kyr at Luochuan and aim to understand the evolution of vegetation and climate change on the Chinese Loess Plateau. The main results and conclusions are as follows: 1. The palynological results show that the grassland has been a dominant vegetation in the Luochuan area since 630 kyr, even during the intervals of relatively warm and wet climatic conditions. 2. The pollen concentration of Luochuan section sharply decreases from the bottom of S1 to downward depth. This decrease can be attributed to depositional environment rather than climate change. In loess, not only oxidation, but also the PH of deposits and bacteria or fungi have been able to degrade sporopollenin. 3. The paleoclimatic condition during S4 stage, characterized with warmer condition during the early stage, was warmer and wetter than that during S5 in Luochuan area. Paleoclimate was warmer and wetter during the early stage of S5 and became colder and drier later. The special pedogenic features of S5-I can be attributed to a prolonged pedogenic duration rather than a warm-wet climate. 4. Evidence from pollen assemblage suggests that the Holocene vegetation has been affected by human impacts, especially after the Yangshao Culture. 5. The present steppe environment on the loess plateau is mainly due to natural conditions. Temperature, seasonal precipitation and soil structure are three important factors which control the vegetation type. 6. The vegetation on the loess plateau is characterized with zonal or azonal distribution. So local conditions should be taken into account when recover natural vegetation. Finally, the restoration and reconstruction of ecosystem on the loess plateau area should be focused on planting grassland rather than forests.

一株岩生真菌对碳酸盐岩的风化作用研究

真菌在矿物和岩石风化过程中起着不可忽视的作用，岩生真菌对岩石的风化作用研究是微生物风化作用研究的热点问题之一。本研究利用从碳酸盐岩表面分离到一株真菌，开展了岩生真菌对碳酸盐岩风化作用的模拟试验，研究了该真菌对不同碳酸盐岩风化作用的过程和效果，分析了真菌代谢产物，发酵液方解石饱和指数以及风化过程中稀土元素的行为，探讨了该真菌对碳酸盐岩风化作用的机理。 对该岩生真菌GZKM01菌株形态学特征和基于ITS序列的分子鉴定结果表明该菌株隶属真菌界(Kingdom fungi)、半知菌亚门(Deuteromycotina)、腔孢纲(Coelomycetes)、球壳孢目(Sphaeropsidales)、鲜壳孢科 (Nectrioidaceae)座壳孢属(Aschersonia)。 实验条件下显示该真菌对碳酸盐岩具有较强的风化作用，可以大大地提高岩石中Ca2+的溶出量，发酵液上清液Ca2+浓度大大高于培养基对照和纯水对照。以加石灰岩矿粉的发酵液(摇床培养)为例，发酵液上清液最高Ca2+浓度分别是死菌对照和纯水对照处理最高Ca2+浓度的3.98倍和13.73倍。 真菌在生长和风化岩石过程中可以产生有机酸、醇类等挥发性代谢产物；真菌矿物相互作用过程中，真菌与矿物形成菌体-矿物复合体或聚集体，真菌胞外分泌物在矿粉表面形成生物膜结构；真菌对碳酸盐岩的风化作用导致发酵液方解石饱和指数SIc不断增大，溶液处于方解石过饱和状态。真菌对碳酸盐岩的风化作用包括有机酸和其他小分子物质的化学降解作用，机械破坏作用，碳酸酐酶的催化作用，真菌的对矿质元素的吸收作用，次生成矿作用等。 真菌作用下，发酵液上清夜稀土元素的含量普遍高于K2HPO4浸泡液中稀土元素的含量, 与K2HPO4浸泡液相比真菌浸出的稀土元素δCe值较大、δEu值较小，富Ce贫Eu，稀土元素分异现象明显；真菌菌体中富Ce贫Eu的特征更加明显，菌体对Ce有强烈的富集作用。代谢产物的络合作用，真菌产生的代谢产物对pH的调节作用，菌体的直接吸收和吸附作用，有机质的吸附作用，氧化还原作用等是造成稀土元素溶出、迁移和富集的主要原因。 分析认为该真菌对碳酸盐岩风化作用包括一系列的生物化学和生物物理过程，这两个过程相互依赖，相互促进，协同作用，岩生真菌在碳酸盐岩的风化成土过程中起着非常重要的作用。该真菌对岩石中稀土元素溶出、迁移和富集同样起着重要作用。通过综合分析风化壳的风化强度、稀土元素分布特征、微生物活动等可以为评估微生物在岩石风化过程中所起的作用提供依据。上述研究加深了对岩生真菌与碳酸盐岩相互作用过程和机理的理解，为进一步研究碳酸盐岩微生物风化作用提供资料。

Controle de fungos associados às sementes de milho por actinomicetos.

2001