Purification and biochemical characterization of feruloyl esterases from Aspergillus terreus MTCC 11096

Aspergillus terreus MTCC 11096 isolated from the soils of agricultural fields cultivating sweet sorghum was previously identified to produce feruloyl esterases (FAEs). The enzymes responsible for feruloyl esterase activity were purified to homogeneity and named as AtFAE‐1, AtFAE‐2, and AtFAE‐3. The enzymes were monomeric having molecular masses of 74, 23 and 36 kDa, respectively. Active protein bands were identified by a developed pH‐dependent zymogram on native PAGE. The three enzymes exhibited variation in pH tolerance ranging between pH 5–8 and thermostability of up to 55°C. Inhibition studies revealed that the serine residue was essential for feruloyl esterase activity; moreover aspartyl and glutamyl residues are not totally involved at the active site. Metal ions such as Ca²⁺, K⁺, and Mg²⁺ stabilized the enzyme activity for all three FAEs. Kinetic data indicated that all three enzymes showed catalytic efficiencies (k_cat/K_m) against different synthesized alkyl and aryl esters indicating their broad substrate specificity. The peptide mass fingerprinting by MALDI/TOF‐MS analysis and enzyme affinity toward methoxy and hydroxy substituents on the benzene ring revealed that the AtFAE‐1 belonged to type A while AtFAE‐2 and AtFAE‐3 were type C FAE. The FAEs could release 65 to 90% of ferulic acid from agrowaste substrates in the presence of xylanase. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:924–932, 2013     

Improving the fermentation performance of saccharomyces cerevisiae by laccase during ethanol production from steam‐exploded wheat straw at high‐substrate loadings

Operating the saccharification and fermentation processes at high‐substrate loadings is a key factor for making ethanol production from lignocellulosic biomass economically viable. However, increasing the substrate loading presents some disadvantages, including a higher concentration of inhibitors (furan derivatives, weak acids, and phenolic compounds) in the media, which negatively affect the fermentation performance. One strategy to eliminate soluble inhibitors is filtering and washing the pretreated material. In this study, it was observed that even if the material was previously washed, inhibitory compounds were released during the enzymatic hydrolysis step. Laccase enzymatic treatment was evaluated as a method to reduce these inhibitory effects. The laccase efficiency was analyzed in a presaccharification and simultaneous saccharification and fermentation process at high‐substrate loadings. Water‐insoluble solids fraction from steam‐exploded wheat straw was used as substrate and Saccharomyces cerevisiae as fermenting microorganism. Laccase supplementation reduced strongly the phenolic content in the media, without affecting weak acids and furan derivatives. This strategy resulted in an improved yeast performance during simultaneous saccharification and fermentation process, increasing significantly ethanol productivity. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013     

Impact of aeration strategy on CHO cell performance during antibody production

Stirred tank bioreactors using suspension adapted mammalian cells are typically used for the production of complex therapeutic proteins. The hydrodynamic conditions experienced by cells within this environment have been shown to directly impact growth, productivity, and product quality and therefore an improved understanding of the cellular response is critical. Here we investigate the sub‐lethal effects of different aeration strategies on Chinese hamster ovary cells during monoclonal antibody production. Two gas delivery systems were employed to study the presence and absence of the air–liquid interface: bubbled direct gas sparging and a non‐bubbled diffusive silicone membrane system. Additionally, the effect of higher gas flow rate in the sparged bioreactor was examined. Both aeration systems were run using chemically defined media with and without the shear protectant Pluronic F‐68 (PF‐68). Cells were unable to grow with direct gas sparging without PF‐68; however, when a silicone membrane aeration system was implemented growth was comparable to the sparged bioreactor with PF‐68, indicating the necessity of shear protectants in the presence of bubbles. The cultures exposed to increased hydrodynamic stress were shown by flow cytometry to have decreased F‐actin intensity within the cytoskeleton and enter apoptosis earlier. This indicates that these conditions elicit a sub‐lethal physiological change in cells that would not be detected by the at‐line assays which are normally implemented during cell culture. These physiological changes only result in a difference in continuous centrifugation performance under high flow rate conditions. Product quality was more strongly affected by culture age than the hydrodynamic conditions tested. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013.     

Chemische Untersuchungen über unterschiedlich vorbehandelte Federmehle

Abstract

Dr. Carlheinrich Engelmann: Ernährung und Fütterung des Geflügels. Erschienen im Neumann-Verlag Radebeul/Berlin, 162 Seiten, 17 × 24 cm Halbleinen, MDN 6,80. Reviewed by H. Bergner

Dr. Martin Pankrath: Wasserbedarf landwirtschaftlicher Nutztiere — Mengennormen. - Erschienen im S. Hirzel Verlag, Leipzig C 1, Schuhmachergäßchen 1–3. 1. Auflage 1963, 99 Seiten mit 25 Diagrammen, 21 Tabellen und 2 Tafeln im Text. 8°, Hln., MDN 10,70. Reviewed by H. Bergner

Physiologische und wirtschaftliche Leistungsgrenzen in der Tierzucht 14 Vorträge eines internationalen Symposions im Max-Planck-Institut für Tierzucht und Tierernährung in Mariensee vom 14. bis 17. Oktober 1963.: Herausgegeben von M. Witt. 315 Seiten mit 8 Kunstdrucktafeln. Englische Summaries. Steif broschiert, Preis DM 16,-. Bestellung über Institut. Reviewed by H. Bergner     

Estimation of net nutrient oxidation and lipogenesis in growing pigs

Data from previous series of trials were reevaluated in order to quantify the net nutrient oxidation and lipogenesis by combining data from balance experiments and indirect calorimetry measurements. The experiments were carried out with eight castrated males of Danish Landrace measured individually from 30 to 100 kg BW. All pigs were fed alternately on high feeding level, near ad libitum and on low feeding level, near maintenance. Oxidation of carbohydrate (OXCHO) and fat (OXF) was calculated from gas exchange measurements. On high feeding level the contribution from OXCHO and OXF to the total heat production was 55 and 30%, respectively, at 30 kg BW and fairly constant from 60 kg BW with OXCHO around 80% and no net OXF. At low feeding level the contribution from OXCHO and OXF was 28 and 63%, respectively, at 30kg BW, increasing to 52% for OXCHO and decreasing to 37% for OXF at 95 kg BW. The lipogenesis on high feeding level increased from 2.4 to 11.9 MJ/d, while on low feeding level it increased from 2.5 to 3.6 MJ/d at 30 to 95 kg BW. The intake of carbohydrate was too low to cover energy requirement by OXCHO on low feeding level and in the first periods on high feeding level and OXF occurred. In spite of a deficit of carbohydrate for oxidation a part of carbohydrate (2.6 MJ/d, ～ 150 g/d) was used for fat synthesis, indicating an importance of lipogenesis in growing pigs.     

Isolation of an aerobic sulfur oxidizer from the SUP05/Arctic96BD-19 clade

Bacteria from the uncultured SUP05/Arctic96BD-19 clade of gamma proteobacterial sulfur oxidizers (GSOs) have the genetic potential to oxidize reduced sulfur and fix carbon in the tissues of clams and mussels, in oxygen minimum zones and throughout the deep ocean (>200 m). Here, we report isolation of the first cultured representative from this GSO clade. Closely related cultures were obtained from surface waters in Puget Sound and from the deep chlorophyll maximum in the North Pacific gyre. Pure cultures grow aerobically on natural seawater media, oxidize sulfur, and reach higher final cell densities when glucose and thiosulfate are added to the media. This suggests that aerobic sulfur oxidation enhances organic carbon utilization in the oceans. The first isolate from the SUP05/Arctic96BD-19 clade was given the provisional taxonomic assignment ‘Candidatus: Thioglobus singularis'', alluding to the clade''s known role in sulfur oxidation and the isolate''s planktonic lifestyle.     

Effects of landscape and management on ground‐dwelling insect assemblages of farmland in Jeju Island,Korea

Granite‐derived soils are widespread in the farmland of Korea in general. In contrast, Jeju Island has mainly volcanic ash soils. Soils and weather condition in Jeju Island created a unique agricultural system. We identified the features of ground‐dwelling insects in farmlands of Jeju Island. This study was conducted in four areas (Samdal‐ri and Susan‐ri in Seogwipo city, and Dongmyeong‐ri and Suwon‐ri in Jeju city) in Jeju Island, Korea. Field surveys were carried out twice in summer (June) and autumn (September) in 2013. Ground‐dwelling insects were sampled quantitatively by using pitfall traps. As a result, in total 3322 individuals, 137 species, 48 families and 8 orders were investigated in farmlands of Jeju Island. Especially, members of Coleoptera and Hymenoptera accounted for a large proportion of ground‐dwelling insect communities. The numbers of species and individuals for major taxonomic groups showed significant regional and seasonal differences. This study implied that the seasonal and regional differences of ground‐dwelling insect communities were affected by surrounding land use patterns, life history patterns of each taxonomic group and farmland management.     

序言Ⅱ 冬虫夏草:从科学到产业

作为名贵传统药材,冬虫夏草的研究方兴未艾。无性型的确定、学名的变更和"一种真菌一个名称"确定了冬虫夏草菌为Ophiocordyceps sinensis;独特的分布、悠久的历史、公众的认可及地方经济的重要作用等被推荐为"国菌";"虫草产业发展金湖宣言"从产、学、研倡议"尊重科学、利好产业"的发展思路。随着科学研究深入和产业技术发展,冬虫夏草培植获得成功。本专刊对冬虫夏草培植相关基础研究和关键技术进行了总结,对寄主昆虫选育及规模化饲养关键技术到冬虫夏草菌菌种筛选、侵染途径和培植以及培植产品与野生冬虫夏草化学成分和蛋白质谱比较等进行了论述。冬虫夏草培植的成功,不论从科学还是产业的角度,都具有里程碑式的意义。     

Fluorescent substrates for ADAM15 useful for assaying and high throughput screening

A disintegrin and metalloproteinase 15 (ADAM15), also known as metargidin, plays important roles in regulating inflammation, wound healing, neovascularization, and is an attractive drug target. Fluorescence resonance energy transfer (FRET)-based peptide substrates were tested to identify candidate reagents for high throughput screening and detection of ADAM15 in biological samples. ADAM15 exhibits a unique and diverse activity profile compared to other metalloproteinases. Two FRET substrates, Dabcyl-Gly-Pro-Leu-Gly-Met-Arg-Gly-Lys(FAM)-NH2 (PEPDAB011) and Dabcyl-Ala-Pro-Arg-Trp-Ile-Gln-Asp-Lys(FAM)-NH2 (PEPDAB017), which also detect activities of several matrix metalloproteinases (MMPs −2, –9, and −13), were efficiently cleaved by ADAM15 with specificity constants of 5800 M⁻¹ s⁻¹ and 4300 M⁻¹ s⁻¹, respectively. Additionally, ADAM15 efficiently processed Dabcyl-Leu-Arg-Glu-Gln-Gln-Arg-Leu-Lys-Ser-Lys(FAM)-NH2 (PEPDAB022), which is based on a physiological CD23 cleavage site, with a specificity constant (k_cat/K_m) of 5200 M⁻¹ s⁻¹. PEPDAB022 was used to screen the ability of known metalloproteinase inhibitors including TAPI-2, marimastat, GI-254023, and the Tissue Inhibitor of Metalloproteinases(TIMPs) 1 and 3 to block ADAM15 activity. Even though ADAM15 exhibits similar substrate preferences to other metalloproteinases, many broad spectrum inhibitors failed to block ADAM15 activity at concentrations as high as 50 μM. Thus, a clear need exists to develop potent and selective ADAM15 inhibitors, and the FRET substrates described herein should aid future research efforts towards this aim.