Cured sweetpotato roots were stored at different temperatures (4.5, 15.6, and 24 degrees C) for 7 weeks and assayed for invertase activities and reducing sugar levels during two separate years. Invertase activities and reducing sugar concentration significantly increased in the roots kept at low temperature. Of the three types of invertases assayed, acid invertase specific activity was the highest. Acid invertase was the most influential in determining reducing sugar levels in stored sweetpotato. Cultivar differences were found in invertase specific activities and reducing sugar concentration. Reducing sugar content was highly correlated to acid and total invertase activity, regardless of cultivar.

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An alternative coupled method was developed for the enzymatic assay of plant invertases. In this method, ADP produced from the phosphorylation of glucose and fructose (hydrolysis products of invertases) is coupled to oxidation of NADH by the enzymes pyruvate kinase and lactate dehydrogenase in the presence of phosphoenolpyruvate and NADH. This method was compared with the glucose-6-phosphate dehydrogenase method for both continuous and discontinuous assay by using purified invertase from baker's yeast and protein preparations derived from plant materials of three different species. This method is applicable to filtered and concentrated crude extracts. Statistical analysis indicated that the alternative method was similar in accuracy to the glucose-6-phosphate dehydrogenase method. Keywords: Biochemistry; enzymes; physiology; sugars; invertase

An appropriate method was developed for the continuous assay of sucrose synthase (SS) (EC 2.4.1.13) by spectrophotometry. The uridine 5'-diphosphate derived from sucrose synthesis was stoichiometrically coupled to oxidation of beta-nicotinamide adenine dinucleotide by the enzymes nucleoside-5'-diphosphate kinase (NDPK), pyruvate kinase, and lactate dehydrogenase. Utilization of crude extracts led to a complete masking of SS assay by adenylate kinase, adenosine 5'-triphosphatase (ATPase), and phosphoenolpyruvate phosphatase found in the crude extracts. These interfering enzymes were mostly removed from the crude extracts by using a combination of gel filtration, centrifugation through a selectively permeable membrane (Biomax-100 Ultrafree centrifugal device), and inhibition by the addition of K(2)HPO(4) to the assay buffer. Sensitivity of the SS assay was significantly increased by the inclusion of NDPK and ATP, which are essential to the reaction in the coupling system.

`Beauregard' sweetpotatoes (Ipomoea batatas L. Lam) were stored under a continuous flow of 0%, 1%, 1.5%, 2%, 5%, 10%, or 21% O2 (balance N2) for 14 days. Respiration rate was significantly lower at 1.5%, 2%, 5%, and 10% O2 compared with 21% O2, while respiration at 0% and 1% O2 was higher than at 1.5%, 2%, 5%, and 10% O2. Respiration rate at 0% O2 remained high for several days after exposure to air while that at 1.5%, 2%, 5%, and 10% O2 increased rapidly to equal that of 21% O2. Ethanol and acetaldehyde accumulated rapidly at 0% and 1% O2 but were lower at the other O2 levels. Ethanol increased 16- and 4-fold after 14 days of storage at 0% and 1% O2, respectively, compared to 21% O2. In addition, acetaldehyde increased 11- and 8-fold at 0% and 1% O2 respectively, compared to 21% O2. Sucrose and total sugar concentration increased under low O2 concentration while reducing sugars (fructose and glucose) and pH decreased.