Microbial and biochemical spoilage of foods: an overview

Biological activity of limonoids from the rutales

Species of the genus Citrus accumulate large quantities of flavanones that affect fruit flavor and have been documented to benefit human health. Bitter species, such as grapefruit and pummelo, accumulate bitter flavanone-7-O-neohesperidosides responsible, in part, for their characteristic taste. Non-bitter species, such as mandarin and orange, accumulate only tasteless flavanone-7-O-rutinosides. The key flavor-determining step of citrus flavanone-glycoside biosynthesis is catalyzed by rhamnosyltransferases; 1,2 rhamnosyltransferases (1,2RhaT) catalyze biosynthesis of the bitter neohesperidosides, while 1,6 rhamnosyltransferases (1,6RhaT) catalyze biosynthesis of the tasteless rutinosides. We report on the isolation and functional characterization of the gene Cm1,2RhaT from pummelo which encodes a citrus 1,2RhaT. Functional analysis of Cm1,2RhaT recombinant enzyme was conducted by biotransformation of the substrates using transgenic plant cell culture. Flavanones and flavones, but not flavonols, were biotransformed into 7-O-neohesperidosides by the transgenic BY2 tobacco cells expressing recombinant Cm1,2RhaT. Immunoblot analysis established that 1,2RhaT protein was expressed only in the bitter citrus species and that 1,6RhaT enzyme, whose activity was previously documented in non-bitter species, was not cross-reactive. Expression of Cm1,2RhaT at the RNA level was prominent in young fruit and leaves, but low in the corresponding mature tissue, thus correlating well with the developmental pattern of accumulation of flavanone-neohesperidosides previously established. Phylogenetic analysis of the flavonoid glycosyltransferase gene family places Cm1,2RhaT on a separate gene cluster together with the only other functionally characterized flavonoid-glucoside rhamnosyltransferase gene, suggesting a common evolutionary origin for rhamnosyltransferases specializing in glycosylation of the sugar moieties of flavonoid glucosides.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-313X.2004.02193.x

Citrus fruit bitter flavors: isolation and functional characterization of the gene Cm1,2RhaT encoding a 1,2 rhamnosyltransferase, a key enzyme in the biosynthesis of the bitter flavonoids of citrus

A panel of 20 to 27 screened judges was used to determine the threshold of pure limonin and naringin in distilled water, sucrose solutions, citric acid solutions adjusted to various pH values and in citrus juice model systems. Thresholds in distilled water were 1 and 20 mg/l for limonin and naringin, respectively. Sucrose, citric acid and combinations of these constituents in a model system increased the threshold of limonin and naringin several fold. The highest thresholds were obtained at low pH values in the absence of sugar. In a 1.2% citric acid solution containing 5% sucrose, 2.5% dextrose and 2.5% fructose the maximum limonin threshold of 11 mg/kg was obtained at pH 3.8. The maximum difference threshold for limonin in orange juice was 6.5 mg/kg and also occurred at pH 3.8. The optimum pH for limonin bitterness suppression (maximum threshold) was 3.8 in both model systems and natural orange juice. Thresholds on either side of this value were lower and limonin bitterness was more noticeable. No pH optimum was observed for naringin threshold in model systems. Individual thresholds showed a wide range of sensitivity to both limonin and naringin bitterness.

Effect of some citrus juice constituents on taste thresholds for limonin and naringin bitterness

Limonoids are heavily oxygenated, modified triterpenes dominant in Meliaceae and Rutaceae plant families. The term 'limonoid' is derived from limonin, which was first identified as the bitter constituent of Citrus seeds in 1841. This group of secondary metabolites exhibits a wide range of biological properties, including anticancer, antibacterial, antifungal, antimalarial, and antiviral activities. Significant progress on the role of limonoids as promising candidates for cancer chemoprevention and/or therapy has been achieved in particular in recent years. The aim of this review article is to discuss the recent developments on limonoids chemical aspects and biological activities with the relationship between structure and activity, supporting the new possibilities for the medicinal and/or nutraceutical use of these compounds.

An overview on chemical aspects and potential health benefits of limonoids and their derivatives.

SUMMARY– Tissues of early-season navel oranges and grapefruit were found by paper electrophoretic procedures to contain a nonbitter precursor of limonin, but no significant amounts of limonin. Limonin is the intensely bitter triterpenoid dilactone responsible for the bitterness which develops in certain navel orange juices on standing. The nonbitter precursor was identified as limonin monolactone by comparison with the authentic compound prepared by partial hydrolysis of limonin and by acid-catalyzed conversion into limonin. Limonin monolactone is stable in the tissues of the intact fruit (which are not bitter) because it is apparently not in direct contact with the acidic juice. It is slowly converted into limonin (and the juice becomes bitter) when the fruit tissues come in contact with the juice, after the juice is expressed from the fruit. Limonin monolactone was not detected in late-season navel oranges or grapefruit. This agrees with the fact that juice made from these fruits does not contain limonin. Work is in progress to determine whether the naturally occurring limonin monolactone is the A-ring monolactone, the D-ring monolactone, or a mixture of the two.

G. D. Beverly

Papers

Limonin Monolactone, the Nonbitter Precursor Responsible for Delayed Bitterness in Certain Citrus Juices