Functional Characteristics of Aldehyde Dehydrogenase and Its Involvement in Aromatic Volatile Biosynthesis in Postharvest Banana Ripening.

Butanol vapor feeding to ripe banana pulp slices produced abundant butyl butanoate, indicating that a portion of butanol molecules was converted to butanoate/butanoyl-CoA via butanal, and further biosynthesized to ester. A similar phenomenon was observed when feeding propanol and pentanol, but was less pronounced when feeding hexanol, 2-methylpropanol and 3-methylbutanol. Enzymes which catalyze the cascade reactions, such as alcohol dehydrogenase (ADH), acetyl-CoA synthetase, and alcohol acetyl transferase, have been well documented. Aldehyde dehydrogenase (ALDH), which is presumed to play a key role in the pathway to convert aldehydes to carboxylic acids, has not been reported yet. The conversion is an oxygen-independent metabolic pathway and is enzyme-catalyzed with nicotinamide adenine dinucleotide (NAD+) as the cofactor. Crude ALDH was extracted from ripe banana pulps, and the interference from ADH was removed by two procedures: (1) washing off elutable proteins which contain 95% of ADH, but only about 40% of ALDH activity, with the remaining ALDH extracted from the pellet residues at the crude ALDH extraction stage; (2) adding an ADH inhibitor in the reaction mixture. The optimum pH of the ALDH was 8.8, and optimum phosphate buffer concentration was higher than 100 mM. High affinity of the enzyme was a straight chain of lower aldehydes except ethanal, while poor affinity was branched chain aldehydes.

Inhibition of acetate ester biosynthesis in banana (Musa sapientum L.) fruit pulp under anaerobic conditions.

The effect of anaerobic conditions on acetate ester biosynthesis in ripened banana pulp was investigated. Incubation of the pulp in less than 1% O(2) resulted in a significant reduction in the formation of ethyl acetate. Regardless of the presence of a large amount of endogenous ethanol and the remaining exogenous isobutyl alcohol after complete anaerobic incubation with the pulp, the production of acetate ester decreased. The effect of addition of pyruvate, isobutyl alcohol, acetate, and methyl hexanoate on acetate ester formation in 100% N(2) was also investigated. The addition of pyruvate and isobutyl alcohol to the pulp gave lower acetate esters in N(2) than in air, whereas the pulp incubated with acetate and isobutyl alcohol produced more acetate ester in both conditions. Therefore, the lack of acetyl CoA, or more precisely acetate, in the tissue is the main reason for the inhibition of acetate ester formation under anaerobic conditions. The activity of beta-oxidation measured by incubation with methyl hexanoate was detected only in the samples incubated in air. The formation of acetyl CoA, derived from pyruvate through mitochondria and through beta-oxidation, was inhibited by anaerobic conditions, which suggests that mitochondrial activity and/or beta-oxidation are essential for ester biosynthesis.