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C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli
(2010)
Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477.
This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7–11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease.
Die SPAC-SE ist börsenfähig und damit eine für eine SPAC grundsätzlich geeignete Rechtsform. Die Tatsache, dass es sich hierbei (zunächst) um eine leere, arbeitnehmerlose Hülle handelt, ändert hieran nichts. Die Gründung einer solchen Vorrats-SE ist trotz fehlender Arbeitnehmerbeteiligung unter teleologischer Reduktion von Art. 12 II SE-VO zulässig. Im Gegenzug muss die Arbeitnehmerbeteiligung gemaß § 18 III SEBG analog nachgeholt werden, wenn das später erworbene Zielunternehmen auf die SPAC-SE verschmolzen werden soll. Aus gesellschaftsrechtlicher Sicht ist zu beachten, dass die SPAC-SE mit Sitz in Deutschland, welche auch den deutschen, aktienrechtlichen Bestimmungen unterliegt, nur bedingt für eine SPAC geeignet erscheint. Das deutsche Aktienrecht enthält strenge Regelungen, die der für eine SPAC-SE erforderlichen Flexibilität entgegenstehen können. Dies gilt insbesondere für das Erfordernis der Zustimmung der Hauptversammlung zur Akquisition des Zielunternehmens, die Rückzahlung des Treuhandvermögens an Aktionäre, die der Akquisition nicht zugestimmt haben und die Liquidation der SPAC-SE im Falle des Scheiterns des Erwerbs des Zielobjektes.
We analyze the trading behavior of individual investors in option-like securities, namely bankissued warrants, and thus expand the growing literature of investors behavior to a new kind of securities. A unique data set from a large German discount broker gives us the opportunity to analyze the trading behavior of 1,454 investors, making 89,958 transactions in 6,724 warrants on 397 underlyings. In different logit regression, we make use of the facts that investors can speculate on rising and falling prices of the underlying with call and put warrants and that we also have information about the stock portfolios of the investors. We report several facts about the trading behavior of individual investors in warrants that are consistent with the literature on the behavior of individual investors in the stock market. The warrant investors buy calls and sell puts if the price of the underlying has decreased over the past trading days and they sell calls and buy puts if the price of the underlying has increased. That means, the investors follow negative feedback trading strategies in all four trading categories observed. In addition, we find strong evidence for the disposition effect for call as well as put warrants, which is reversed in December. The trading behavior is also influenced if the underlying reaches some exceptionally prices, e.g. highs, lows or the strike price. We show that hedging, as one natural candidate to buy puts, does not play an important role in the market for bank-issued warrants.
Bussysteme
(2006)