Simultaneous saccharification and ethanol fermentation at high corn stover solids loading in a helical stirring bioreactor

The higher ethanol titer inevitably requires higher solids loading during the simultaneous enzymatic saccharification and fermentation (SSF) using lignocellulose as the feedstock. The mixing between the solid lignocellulose and the liquid enzyme is crucially important. In this study, a bioreactor with a novel helical impeller was designed and applied to the SSF operation of the steam explosion pretreated corn stover under different solids loadings and different enzyme dosages. The performances using the helical impeller and the common Rushton impeller were compared and analyzed by measuring rheological properties and the mixing energy consumption. The results showed that the new designed stirring system had better performances in the saccharification yield, ethanol titer, and energy cost than those of the Rushton impeller stirring. The mixing energy consumption under different solids loadings and enzyme dosages during SSF operation were analyzed and compared to the thermal energy in the ethanol produced. A balance for achieving the optimal energy cost between the increased mixing energy cost and the reduced distillation energy cost at the high solids loading should be made. The potentials of the new bioreactor were tested under various SSF conditions for obtaining optimal ethanol yield and titer. Biotechnol. Bioeng. 2010. 105: 718–728. © 2009 Wiley Periodicals, Inc.     

悬铃木方翅网蝽: 一种正在迅速扩张的城市外来入侵害虫

悬铃木方翅网蝽(Corythucha ciliata)是新近入侵我国城市生态系统的外来害虫, 严重危害城市行道树悬铃木(Platanus spp.), 自2002年该虫在湖南长沙首度报道以来, 已在我国11个省(直辖市)的25个城市发生, 尤其在长江流域呈现爆发态势。本文系统介绍了悬铃木方翅网蝽的生活史、生活习性、繁殖潜力等生物学特性, 以及扩散速度与方式、影响发生的主要环境因素、种群的耐寒耐热能力等生态学特性。最后, 在分析该虫的寄主及危害特点的基础上, 对其控制与管理对策进行了综述, 以为其管理提供参考。     

A novel metagenome-derived β-galactosidase: gene cloning, overexpression, purification and characterization

A novel β-galactosidase gene, zd410, was isolated by screening a soil metagenomic library. Sequence analysis revealed that zd410 encodes a protein of 672 amino acids with a predicted molecular weight of 78.6 kDa. The recombinant ZD410 was expressed and purified in Pichia pastoris, with a yield of ca. 300 mg from 1 L culture. The purified enzyme displayed optimal activity at 38°C and pH 7.0. Given that the enzyme had 54% of the maximal activity at 20°C and 11% of the maximal activity at close to 0°C, ZD410 was regarded as a cold-adapted β-galactosidase. ZD410 displays high enzymatic activity for its synthetic substrate-ONPG (o-nitrophenyl-β-d-galactopyranoside, 243 U/mg) and its natural substrate-lactose (25.4 U/mg), while its activity was slightly stimulated by addition of Na⁺, K⁺, or Ca²⁺ at low concentrations. ZD410 is a good candidate of β-galactosidases for food industry after further study.     

北细辛内生真菌的分离鉴定及代谢产物的生物活性

采用表面消毒法分别从3种北细辛中分离获得10株形态特征不同的优势内生真菌,经形态学和18S rDNA ITS分子分类学分析鉴定为小丛壳属(Glomerella sp.)、叶点霉属(Phyllosticta sp.)、柄孢壳菌属(Po-dospora sp.)、刺盘孢属(Colletotrichum sp.)和镰孢属(Fusarium sp.)。对北细辛优势内生真菌的发酵产物进行体外抗肿瘤和抗菌活性检测,结果表明:除菌株E3、E4和E10外,其余菌株均有不同程度的抗肿瘤和抗菌活性;镰孢属(Fusarium sp.)菌株E9对A549、MDA-MB-231和PANC-1肿瘤细胞抑制率达75%以上;小丛壳属(Glomerella sp.)菌株E1和叶点霉属(Phyllosticta sp.)菌株E2靶向FabI的抗菌活性较强,抑制率达59%。     

Suppression of CD4 T-Cells in the spleen of mice infected with Toxoplasma gondii KI-1 tachyzoites

Toxoplasma gondii KI-1, a recent new isolate from Korea, shows similar pathogenicity and infectivity to mice compared to the virulent RH strain. To understand characteristics of host immunity, including immune enhancement or suppression, we investigated proliferative responses and phenotypes of spleen cells. In addition, kinetics of IFN-γ, a Th1 cytokine, was examined in BALB/c mice up to day 6 post-infection (PI). Intraperitoneal injection of mice with 10(3) KI-1 tachyzoites induced significant decreases (P < 0.05) in proliferative responses of spleen cells. This occurred at days 2-6 PI even when concanavalin A (con A) was added and when stimulated with KI-1 antigen, suggesting suppression of the immunity. CD4(+) T-cells decreased markedly at day 2 PI (P < 0.05), whereas CD8(+) T-cells, NK cells, and macrophages did not show significant changes, except a slight, but significant, increase of CD8(+) T-cells at day 6 PI. The capacity of splenocytes to produce IFN-γ by con A stimulation dropped significantly at days 2-6 PI. These results demonstrate that intraperitoneal injection of KI-1 tachyzoites can induce immunosuppression during the early stage of infection, as revealed by the decrease of CD4(+) T-cells and IFN-γ.     

Antisense-mediated Depletion of Tomato Endoplasmic Reticulum Omega-3 Fatty Acid Desaturase Enhances Thermal Tolerance

An endoplasmic reticulum-localized tomato ω-3 fatty acid desaturase gene(LeFAD3) was isolated.The antisense tomato plants were obtained under the control of the cauliflower mosaic virus 35S promoter(35S-CaMV).Northern blot analysis confirmed that the expression of LeFAD3 was inhibited in the tomato genome.Levels of 18:3 decreased and correspondingly levels of 18:2 increased in total lipids of leaves and roots.After heat stress,the fresh weight of the aerial parts of antisense transgenic plants was higher than that of the wild type(WT) plants.The membrane system ultrastructure of chloroplasts in leaf cells and all of the subcellular organelles in the root tips of transgenic plants remained more intact than those of WT.Relative electric conductivity increased less in transgenic plants than in WT.Under heat stress,the maximal photochemical efficiency of photosystem II(Fv/Fm) and the O2 evolution rate decreased more in WT than in transgenic plants.These results suggested that the depletion of LeFAD3 increased the saturation of fatty acids and alleviated high temperature stress.     

The influence of water salinity and stage in life history on ion concentration of fish blood serum

(1) The Na, K, Ca, Mg concentration of blood serum have been studied in nearly 60 fish species from various waterbodies (the Neva, Lake Dal'nee, the Volga, Lake Baikal, the White Sea, the Black Sea, the Barents Sea etc.). In fishes (Cyprinidae, Percidae, Siluridae etc.) from fresh water (Lake Dal'nee, the Baikal, the Neva) with low 0.18–0.24 mEq/1 Na concentration Na content in blood serum was lower than in closely related species from fresh water (the Volga, Lyutoga River) with larger Na concentration–nearly 0.5–1 mEq 1. Blood serum of marine teleosts Na concentration (156–210 mEq/1) was higher than in migrating and freshwater species (90–174) mEq/1) and it did not depend on seawater salinity: Barents Sea–465, White Sea–255, Black Sea–227 mEq/1.
(2) Rather a considerable specific variation of ion concentration was found in blood serum of fishes from different families living in the same water body (Lake Baikal: Cyprinidae 90–95 mEq/1, Percidae 114 mEq/1, Salmonidae 126–129 mEq/1 etc.; the Volga: Siluridae 139 mEq/1, Percidae 142–150 mEq/1, Clupeidae 174 mEq/1).
(3) No relation has been detected between K, Ca and Mg concentrations in blood serum of fishes and the concentrations of these ions in the environment.
(4) Na concentration in the blood serum of sockeye salmon juveniles (101 mEq/1) migrating seaward was less than in adult fishes (132 mEq/1) returning to the lake to spawn. Na concentration in the blood of non-migrating salmonids ( Oncorhynchus nerka adult residual 104 mEq/1, Salvelinus alpinus juvenile 140 mEq/1, adult residual 141 mEq/1) changed insig- nificantly during life history. Notable hypopotassemia was observed in spawning time in Salmonidae.
(5) The comparison of blood serum ion concentration in males and females of fishes from different taxonomic groups (Acipenseridae, Salmonidae, Cyprinidae) showed no significant variation.     

NMR Structural Studies of Domain 1 of Receptor-associated Protein

The 39 kDa receptor-associated protein (RAP) is an endoplasmic reticulum resident protein that binds tightly to the low-density lipoprotein receptor-related protein (LRP) as well as to other members of the low-density lipoprotein receptor superfamily. The association of RAP with LRP prevents this receptor from interacting with ligands. RAP is a three-domain protein that contains two independent LRP binding sites; one located within domains 1 and 2, and one located within domain 3. As the first step toward defining the structure of the full-length protein and understanding the interaction between RAP and this family of receptors, we have determined the 3D structure of domain 1 using constraints derived from heteronuclear multi-dimensional NMR spectra, including NOEs, dihedral angles, J-couplings and chemical shifts, as well as two sets of non-correlated residual dipolar couplings measured from the protein solutions in anisotropic media of Pf1 and 6% polyacrylamide gel. The backbone C(alpha) rmsd between the current structure and a homo-nuclear NOE-based structure is about 2 A. The large rmsd mainly reflects the significant differences in helical orientation and in the structural details of the long helix (helix 2) between the two structures.     

Enzymatic activity of glucose oxidase covalently wired via viologen to electrically conductive polypyrrole films

The surface functionalization of an electrically conductive polypyrrole film (PPY) with a viologen, (N-(2-carboxyl-ethyl)-N'-(4-vinyl-benzyl)-4,4'-bipyridinium dichloride, or CVV) for the covalent immobilization of glucose oxidase (GOD) has been carried out. The viologen was first synthesized and graft polymerized on PPY film. It then served as an anchor via its carboxyl groups for the covalent immobilization of GOD. The surface composition of the as-functionalized substrates was characterized by X-ray photoelectron spectroscopy (XPS). The effects of the CVV monomer concentration on the CVV-graft polymer concentration and the amount of GOD immobilized on the surface were investigated. The activity of the immobilized GOD was compared with that of free GOD and the kinetic effects were also obtained. The cyclic voltammetric (CV) response of the GOD-functionalized PPY substrates was studied in a phosphate buffer solution under an argon atmosphere. The CV results support the mechanism in which CVV acts as a mediator to transfer electron between the electrode and enzyme, and hence regenerating the enzyme in the enzymatic reaction with glucose. High sensitivity and linear response of the enzyme electrode was observed with glucose concentration ranging from 0 to 20 mM.