| Quantitative Determination of Ascorbate from the Green Alga Chlamydomonas reinhardtii by HPLC
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Author:
Date:
2016-12-20
[Abstract] Ascorbate (Asc, also called vitamin C) is of vital importance to the cellular functions of both animals and plants. During evolution, Asc has become one of the most abundant metabolites in seed plants; however, Asc contents in cyanobacteria, green algae and bryophytes are very low. Here we describe a sensitive and reliable HPLC method for the quantitative determination of cellular Asc content in the green alga Chlamydomonas reinhardtii.
[摘要] 抗坏血酸（Asc，也称为维生素C）对动物和植物的细胞功能至关重要。 在进化过程中，Asc已经成为种子植物中最丰富的代谢物之一; 然而，蓝藻，绿藻和苔藓植物中的Asc含量非常低。 在这里，我们描述了一种灵敏可靠的HPLC方法，用于定量测定绿藻衣藻中细胞Asc含量。
背景 针对高等植物开发了用于确定细胞Asc含量的先前方案，其中细胞Asc的量足够高以用于分析。 这些方案不适用于绿色藻类，因为植物材料的量和细胞Asc的水平通常都受到限制。 因此，需要开发一种灵敏度提高到μM范围来测量绿藻中细胞Asc的新方法。
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| Measurement of Glucose-6-phosphate Dehydrogenase Activity in Bacterial Cell-free Extracts
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Author:
Date:
2016-10-05
[Abstract] Glucose-6-phosphate dehydrogenase (G6PDH) (EC 1.1.1.49) is the first enzyme of the oxidative pentose phosphate cycle and catalyses the conversion of glucose-6-phosphate (G6P) to 6-phosphoglucono-δ-lactone and transfers one electron to NADP+ producing one NADPH. Conversion of G6P to 6-phosphoglucono-δ-lactone is proportional to the production of NADPH. The increase in NADPH concentration results in an increase in absorbance at 340 nm. To assay G6PDH activity, therefore, production of NADPH is determined by measuring increase in absorbance at 340 nm spectrophotometrically. This ...
[摘要] 葡萄糖-6-磷酸脱氢酶（G6PDH）（EC 1.1.1.49）是氧化戊糖磷酸循环的第一个酶，并催化葡萄糖-6-磷酸（G6P）转化为6-磷酸葡萄糖酸-δ-内酯并将一个电子 到NADP +产生一种NADPH。 G6P向6-磷酸葡萄糖酸-δ-内酯的转化与NADPH的产生成比例。 NADPH浓度的增加导致340nm处吸光度的增加。 因此，为了测定G6PDH活性，通过分光光度法测量340nm处吸光度的增加来确定NADPH的产生。 然后将该增加速率转化为G6PDH的活性和比活性的单位。 在该程序中，给出了强调蓝细菌集胞藻的细菌G6PDH测定的一般方法。 PCC6803（Schaeffer和Stanier，1978; Karakaya等人，2008，2012）和异养细菌大肠杆菌（Hylemon和Phibbs，1972; Barnel等人 ，1990）。
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