Influence of pH on recombinant human growth hormone production by Pichia pastoris

BACKGROUND: Effect of pH on recombinant human growth hormone (rhGH) production by Pichia pastoris hGH‐Mut⁺ was investigated at pH = 4.2, 5.0, 5.5, and 6.0. RESULTS: The highest cell concentration was obtained at pH = 6.0 with 53 g L^?1, while the highest rhGH concentration was attained at pH = 5.0 as 0.27 g L^?1. Total protease secretion increased with increase in pH and with the cultivation time. Oxygen uptake rate increased with increasing pH up to pH = 6.0, having the maximum value, 37 mmol m^?3 s^?1, at pH = 5.5. K_La values were similar at all the conditions, having a maximum value of 0.14 s^?1 at pH = 5.0. Taking the final rhGH concentration into account, the most favourable pH was 5.0; where AOX1 expression level showed a similar trend to AOX activity profiles, having the highest value of 9.4 × 10¹⁰ copy mg^?1 CDW at t = 15 h; in parallel to AOX1 expression profile, hGH expression level increased until t = 15 h, with the highest value of 4.0 × 10¹⁰ copy mg^?1 CDW, where a sharp increase in rhGH concentration was obtained. The expression levels of pep4, prb1 and prc1 genes, responsible from the production of proteinase A, proteinase B and, carboxypeptidase Y, were parallel to each other. CONCLUSION: Since it was shown that pH is a crucial operating parameter in fermentation processes using P. pastoris, keeping pH constant at its determined optimum value, pH = 5.0, during the bioprocess is vital in terms of recombinant protein production. Copyright © 2010 Society of Chemical Industry     

木聚糖酶预处理降低漂白废水中的AOX

研究了木聚糖酶预处理对后续漂白废水中AOX含量的影响.研究表明,木聚糖酶预处理降低了纸浆中的木素含量,提高了后续漂白中纸浆的可漂性,后续漂白中在不影响纸浆白度的情况下,降低漂剂用量,减少漂后废水中AOX的含量.     

竹浆木聚糖酶预处理对常规CEH三段漂的影响

进行了竹浆木聚糖酶预处理对常规CEH三段漂影响的研究.实验结果表明:在总用氯量均为8.5%时,用木聚糖酶预处理的硫酸盐竹浆的白度增加了1.99%ISO;在用氯量减少了11.76%的条件下,酶预处理竹浆能够达到普通CEH三段漂的白度,同时,纸浆的裂断长、撕裂度和耐折度分别提高了7%、11%和32%;此外,用木聚糖酶预处理后再漂白工艺可有效降低废水污染负荷.     

二噁英对人类健康的危害及造纸工业的对策

二噁英是强致癌物质,其在环境中的半衰期长达10～20年,是危害性很大的环境激素,可通过食物链进入人体,在动物和人的脂肪中蓄积,对人体健康构成严重威胁.本文介绍了二噁英的命名、结构、性质、来源以及对人体健康的影响,同时介绍了制浆造纸工业控制和减少二噁英的技术措施.     

我国制浆造纸AOX的来源分析及其减量化建议

使用AOX分析仪分析了若干纸张、纸浆、制浆原料和相关化学品AOX的含量,初步了解了制浆造纸AOX的来源并提出了几点AOX减量化建议。结果表明,收集到的纸张、纸浆、化学品、原料等都含有AOX。制浆造纸AOX的来源主要有原料本身的农药残留、水源或者化学品以及含氯漂白剂的使用。可以采用源头控制、清洁生产技术以及制定相关产品标准来减少我国制浆造纸AOX的产生量。     

Analysis of methanol–ethanol mixtures from falsified beverages using a dual biosensors amperometric system based on alcohol dehydrogenase and alcohol oxidase

A portable bienzymatic analytical system was developed for the chronoamperometric analysis of methanol–ethanol mixtures. The system consists of two biosensors, one based on alcohol dehydrogenase (ADH) that responds only to the ethanol and the second one based on alcohol oxidase (AOX) that responds to both methanol and ethanol. The transducers were screen-printed electrodes (SPEs) modified with mediators: Meldola blue for ADH and Co-phthalocyanine for AOX. The calibration graph of the ADH biosensor is linear between 0.3 and 8 mmol/L ethanol. The AOX biosensor is able to quantify both analytes in mixtures that contain methanol between 3 and 70 mmol/L and ethanol ranging from 15 to 110 mmol/L. Interferences due to non-specific oxidations from common oxidizable compounds like gallic acid and ascorbic acid were smaller in the case of transducer based on Meldola blue. The analytical system was successfully tested on real samples: non-alcoholised beer (NAB) spiked with ethanol or methanol and a falsified rose wine (FRW).     

无污染漂白--造纸工业可持续发展的要求

分析了造纸业在可持续发展中的重要地位，并从可持续发展角度论述了纸浆无污染漂白的重要性和必要性，指出无污染漂白是解决制浆工业面临的化工药品浪费大、污染严重等问题的有效途径，并提出了发展无污染漂白技术的对策。     

Organochlorine Formation in Tertiary Amine Catalyzed Pulp Bleaching

An oxygen delignified eucalyptus kraft pulp was bleached by applying a catalytic amount of a tertiary amine, 1,4-diazabicyclo[2,2,2]octane, and hypochlorous acid at varying pH, temperature, and time. The bleaching filtrate was analyzed for chloroform and AOX and the pulp for residual lignin, hexenuronic acid, and OX. In comparison with chlorine dioxide bleaching, the novel catalytic bleaching (pH < 6) generated only traces of chloroform and low contents of AOX and OX. Separate catalytic oxidations of authentic hexenuronic acid showed that it was a major precursor of chloroform. The results demonstrate a practical way to reduce organochlorine formation in pulp bleaching and provide new understanding on the reaction pathways that yield chloroform.     

Lower Temperature Cultures Enlarge the Effects of Vitreoscilla Hemoglobin Expression on Recombinant Pichia pastoris

An heterologous expression of Vitreoscilla hemoglobin (VHb) for improving cell growth and recombinant protein production has been successfully demonstrated in various hosts, including Pichia pastoris. Lower temperature cultures can enhance target protein production in some studies of P. pastoris. In this study, the strategy of combining heterologous VHb expression and lower temperature cultures in P. pastoris showed that final cell density and viability of VHb⁺ strain at 23 °C were higher than that at 30 °C. In addition, the effects of VHb expression on recombinant β-galactosidase production and oxygen uptake rate were also higher at 23 °C than at 30 °C. Consequently, lower temperature cultures can enlarge VHb effectiveness on cell performance of P. pastoris. This is because VHb activity obtained at 23 °C cultures was twofold higher than that at 30 °C cultures, due to a different heme production. This strategy makes P. pastoris an excellent expression host particularly suitable for increasing the yields of the low-stability and aggregation-prone recombinant proteins.     

ECF漂白过程中AOX减量技术研究进展

对纸浆漂白过程中可吸附有机卤素(AOX)减量技术的研究背景与国内外研究现状进行了论述;并通过调查ECF漂白过程中AOX减量技术的研究进展,对AOX相关减量技术做出如下概括：①优化工艺;通过调整漂白工序减少含氯漂剂的用量,从而减少AOX的生成量。②添加化学助剂;通过添加H2O2或添加含硫基、氮基等助剂抑制AOX产生。③漂前预处理;纸浆漂白之前采用热水预抽提、酸碱预处理以及生物酶预处理等方法处理纸浆,减少纸浆中的木素和半纤维素含量,从而降低AOX生成量。