Decreased production ofpara-hydroxypenicillin V in penicillin V fermentations

Summary Penicillin V (phenoxymethyl penicillin) is produced by industrial strains ofPenicillium chrysogenum in the presence of phenoxyacetic acid (POAc), a side-chain precursor for the penicillin V molecule. The wild-type strain ofP. chrysogenum produces an undesirable penicillin byproduct,para-hydroxypenicillin V (p-OH penicillin V), in addition to penicillin V, viapara-hydroxylation of POAc and subsequent incorporation of thep-OH phenoxyacetic acid into the penicillin molecule. Most of thep-OH penicillin V is produced late in cycle when the POAc concentration in the medium is nearly depleted. The level ofp-OH penicillin V produced by the control strain ranges up to 10–15% of the total penicillins produced. 3-Phenoxypropionic acid andp-bromophenylacetic acid partially inhibit the formation ofp-OH penicillin V with a minimal effect on penicillin V productivity. Mutants deficient in their ability to hydroxylate POAc were found to produce lower levels ofp-OH penicillin V. Multi-step mutation and screening, starting with the wild-type strain, have culminated in isolation of mutants which producep-OH penicillin V as 1% of the total penicillins with no adverse effect on penicillin V productivity.     

Multiobjective optimization of the antioxidant activities of horse mackerel hydrolysates produced with protease mixtures

Fish protein hydrolysates (FPH) from horse mackerel were produced by employing an enzyme mixture of subtilisin and trypsin. The antioxidant activity of fish hydrolysates (DPPH scavenging activity, Fe²⁺ chelating activity and Fe³⁺ reducing power) was modelled as a function of the operating conditions for the hydrolysis (i.e. protein concentration, temperature and composition of the enzyme mixture). The antioxidant activities showed different behavior depending on whether their controlling pathway was the transference of electrons/protons (i.e. DPPH scavenging activity and Fe³⁺ reducing power) or metal chelation. In the first case, the antioxidant activities increased with the decrease of substrate concentration and temperature when pure trypsin (DPPH scavenging activity) or a mixture of enzymes (Fe³⁺ reducing capacity) was employed. Contrarily, hydrolysates showed higher Fe²⁺ chelating activities at moderate concentration and high temperature (i.e. 5 g/L and 55 °C) employing solely subtilisin. The conflictive behavior among the antioxidant properties suggested using a multiobjective optimization technique. The ε-constraint method was chosen for this purpose. This approach allows determining the most adequate operational conditions for producing hydrolysate with a specific antioxidant profile which is the first approximation to the production of taylor-made antioxidant hydrolysates.     

Adaptation as organism design

The problem of adaptation is to explain the apparent design of organisms. Darwin solved this problem with the theory of natural selection. However, population geneticists, whose responsibility it is to formalize evolutionary theory, have long neglected the link between natural selection and organismal design. Here, I review the major historical developments in theory of organismal adaptation, clarifying what adaptation is and what it is not, and I point out future avenues for research.     

半定规划鲁棒不可行性研究

本文给出了一种判别半定规划鲁棒不可行的准则,通过该准则我们可判断模型对输入数据是否敏感,进而决定是否应该利用鲁棒优化建模的方法对模型进行重新建模,使其所求解具有鲁棒性.避免了所给模型在实际应用中存在较大的风险性,为更好的应用鲁棒优化建模方法提供了理论依据.     

Effects of temperature and glycerol and methanol‐feeding profiles on the production of recombinant galactose oxidase in Pichia pastoris

Optimization of protein production from methanol‐induced Pichia pastoris cultures is necessary to ensure high productivity rates and high yields of recombinant proteins. We investigated the effects of temperature and different linear or exponential methanol‐feeding rates on the production of recombinant Fusarium graminearum galactose oxidase (EC 1.1.3.9) in a P. pastoris Mut⁺ strain, under regulation of the AOX1 promoter. We found that low exponential methanol feeding led to 1.5‐fold higher volumetric productivity compared to high exponential feeding rates. The duration of glycerol feeding did not affect the subsequent product yield, but longer glycerol feeding led to higher initial biomass concentration, which would reduce the oxygen demand and generate less heat during induction. A linear and a low exponential feeding profile led to productivities in the same range, but the latter was characterized by intense fluctuations in the titers of galactose oxidase and total protein. An exponential feeding profile that has been adapted to the apparent biomass concentration results in more stable cultures, but the concentration of recombinant protein is in the same range as when constant methanol feeding is employed. © 2014 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 30:728–735, 2014     

An improved neutral landscape model for recreating real landscapes and generating landscape series for spatial ecological simulations

Many studies have assessed the effect of landscape patterns on spatial ecological processes by simulating these processes in computer‐generated landscapes with varying composition and configuration. To generate such landscapes, various neutral landscape models have been developed. However, the limited set of landscape‐level pattern variables included in these models is often inadequate to generate landscapes that reflect real landscapes. In order to achieve more flexibility and variability in the generated landscapes patterns, a more complete set of class‐ and patch‐level pattern variables should be implemented in these models. These enhancements have been implemented in Landscape Generator (LG), which is a software that uses optimization algorithms to generate landscapes that match user‐defined target values. Developed for participatory spatial planning at small scale, we enhanced the usability of LG and demonstrated how it can be used for larger scale ecological studies. First, we used LG to recreate landscape patterns from a real landscape (i.e., a mountainous region in Switzerland). Second, we generated landscape series with incrementally changing pattern variables, which could be used in ecological simulation studies. We found that LG was able to recreate landscape patterns that approximate those of real landscapes. Furthermore, we successfully generated landscape series that would not have been possible with traditional neutral landscape models. LG is a promising novel approach for generating neutral landscapes and enables testing of new hypotheses regarding the influence of landscape patterns on ecological processes. LG is freely available online.     

海洋生物技术在海水养殖动植物品种培育和病害防治中的应用

海水养殖是我国沿海地区发展经济的重要支柱产业。目前影响我国海水养殖业持续、稳定、健康发展的主要制约因素是苗种和病害问题。本文就海洋生物技术在海水养殖物植物品种培育和病害防治中的应用及有关问题进行了探讨,分别就应用海洋生物技术在群体和个体水平、细胞和染色体水平以及基因和分子水平培育新品种的工作,包括ＳＰＦ和ＳＰＲ苗种的培育、性别控制和杂交育种、多倍体育种、雌核发育、藻类细胞工程育种、转基因技术等进行了讨论。在重要水产养殖疾病病原的快速诊断和检测技术方面,重卢、讨论了核酸探针技术,ＰＣＲ技术和单克隆抗体技术。海洋生物技术在水产养殖疾病防治中的应用,重点讨论了疫苗和免疫促进剂的开发以及基因工程疫苗。本文还对我国海洋生物技术的发展战略及有关问题进行了探讨。     

A high-throughput media design approach for high performance mammalian fed-batch cultures

An innovative high-throughput medium development method based on media blending was successfully used to improve the performance of a Chinese hamster ovary fed-batch medium in shaking 96-deepwell plates. Starting from a proprietary chemically-defined medium, 16 formulations testing 43 of 47 components at 3 different levels were designed. Media blending was performed following a custom-made mixture design of experiments considering binary blends, resulting in 376 different blends that were tested during both cell expansion and fed-batch production phases in one single experiment. Three approaches were chosen to provide the best output of the large amount of data obtained. A simple ranking of conditions was first used as a quick approach to select new formulations with promising features. Then, prediction of the best mixes was done to maximize both growth and titer using the Design Expert software. Finally, a multivariate analysis enabled identification of individual potential critical components for further optimization. Applying this high-throughput method on a fed-batch, rather than on a simple batch, process opens new perspectives for medium and feed development that enables identification of an optimized process in a short time frame.