Chen-Hsien Lee

Betalains, phase II enzyme-inducing components from red beetroot (Beta vulgaris L.) extracts.

Abstract: Crude aqueous and ethanolic extracts of root tissue of red (Rd) and high-pigment (HP) beet (Beta vulgaris L.) strains exhibited antioxidant and phase II enzyme-inducing activities, and these extracts were fractionated using Sephadex LH-20 chromatography. These bioactivities tended to become co-enriched in early and late eluting fractions, comprising 5-25% of the material recovered from the column. Liquid chromatography-mass spectrometry (MS) was used to resolve and identify multiple betalain components in the most potent quinone reductase (QR)-inducing fractions. Active fractions were found to contain vulgaxanthins I and II, and (iso)betanin, but other components remained unidentified. Two of the isolated active fractions were incorporated into rodent diets at 10-150 ppm over a 2-mo period to assess bioavailability and in vivo efficacy for phase II enzyme induction in various organs. No statistically significant effect of diet was obtained, and wide ranges of tissue enzyme levels among individual animals were observed. This lack of effect and diversity in response to diet may be related to the wide range in absorptive capacity of and/or insufficient level or enrichment of the active agents or to difficulties in assessing such activity in vivo. Subsequent to the animal studies, betanin was isolated in pure form, identified by MS analysis, and confirmed to be QR inducers in the bioassay.

Comparative selectivities of immobilized lipases from Pseudomonas cepacia and Candida antarctica (fraction B) for esterification reactions with glycerol and glycerol analogues in organic media

Abstract Esterification reaction selectivity in homogeneous organic media was evaluated for Candida antarctica B and Pseudomonas cepacia lipases toward members of an homologous series of even carbon number, saturated n -fatty acids (FAs) of 4–16 carbons (C4–C16), glycerol, and glycerol analogues ( n -propanol, isopropanol, 1,2-, and 1,3-propanediols). Dual FA optima were observed in almost all reaction mixtures, with C. antarctica and P. cepacia lipases generally favoring C10/C6 and C8/C16, respectively. Several explanations are offered to account for multiple FA optima. Reaction selectivity toward FAs was more influenced by steric than electronic features of FA substrates. Relative selectivity and discriminatory power among the series of FAs was modulated by the specific alcohol acceptor used. This may be founded on partial sharing of binding pockets by alcohol and FA co-substrates. Evidence was obtained that implicates the sn -2 alcohol functional group as being a determinant of FA selectivity and lipase discriminatory power.

Comparative Fatty Acid Selectivity of Lipases in Esterification Reactions with Glycerol and Diol Analogues in Organic Media

Reaction selectivity of Pseudomonas cepacia, Rhizomucor miehei, and Candida antarctica B lipases was assessed in multicompetitive esterification reaction mixtures containing an homologous series of n‐chain even carbon number fatty acid (FA; C4−C18) substrates and a single alcohol cosubstrate (glycerol, 1,2‐propanediol (1,2‐PD), or 1,3‐propanediol (1,3‐PD)) in tert‐butyl methyl ether at water activity of 0.69 or 0.90 and a reaction temperature of 35 °C. For P. cepacia lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C10, C6, C16 > other FA; with 1,2‐PD and 1,3‐PD, C16 > C8 > C14 > other FA. For R. miehei lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C12 > C10, C14 > other FA; with 1,2‐PD and 1,3‐PD, C8 > C12 > other FA. For C. antarctica B lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C10, C6, C12 > other FA; with 1,2‐PD, C8 > C10, C6 > other FA; and with 1,3‐PD, C8 > C10 > C6 > other FA. The differences in selectivity among FA ranged up to 16‐fold, depending upon the lipase and alcohol cosubstrate used. These findings represent intrinsic and substrate‐modulated features of FA selectivities that are of particular relevance to the use of lipases for acylglycerol synthesis reactions.

Cysteine and Glutathione Mixed-Disulfide Conjugates of Thiosulfinates: Chemical Synthesis and Biological Activities

The chemical syntheses of cysteine (CYS) and glutathione (GSH) mixed -disulfide conjugates (CySSR, GSSR, respectively) of mercapto residues representing most of the R groups of thiosulfinates (R = methyl, ethyl, propyl, and allyl) are described. Gram-scale conjugates were prepared as >98% pure preparations, with 80% reaction yield for each of the two seminal synthesis steps, with structures confirmed by (1)H NMR and high-resolution MS analyses. These conjugates are derivatives of thiosulfinates that may be evolved in processed foods, in the digestive tract, and through in vivo metabolism. The prepared conjugates were found to be able to induce quinone reductase (QR, a representative phase II enzyme) in murine hepatoma cells (Hepa 1c1c7) and to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophage cells (RAW 264.7), indicating they have potential cancer preventive and anti-inflammatory activities. Among the prepared conjugates, the allyl conjugates of CYS and GSH, S-allylmercaptocysteine (CySSA) and S-allylmercaptoglutathione (GSSA), showed the most potent activity regarding QR induction and NO production inhibition. The conjugates with saturated R groups were also active and conferred biological activity as cystine and oxidized glutathione exhibited no effects in these cellular assays.

Relationship between thiosulfinates and pink discoloration in onion extracts, as influenced by pH

Abstract The influence of immediate and delayed pH control, and added S-methyl-L-cysteine sulfoxide (MCSO) on the progress of pink discoloration in relation to the fate of thiosulfinates in extracts of yellow onion (Allium cepa) bulbs was evaluated over an incubation period of about 3 weeks at 21–24°C. Greatest discoloration occurred in extracts prepared with immediate pH control at 6.1, and least discoloration occurred at pH 3.2, over the pH range of 3.2–6.1. Initial thiosulfinate levels, in descending order of abundance, occurred at pH 6.1 > 5.0 > 4.2 > 3.2 for extracts subject to immediate pH control. However, over 21 days incubation, the lowest residual levels (ranging 20–60% of maximum levels) were observed for extracts adjusted to pH 3.2 and 6.1. Extracts subject to delayed (10 min after tissue disruption) pH control (initial extract pH of 5.2–5.4) displayed the greatest extent of discoloration, in descending order, at pH 4.0 > 5.0, 6.0 > 3.0. In these samples, initial thiosulfinate levels were similar and the greatest residual thiosulfinate levels over the 21-day incubation period were observed at pH 3.0 and 4.0 (about 50% of maximum levels), with the lowest residual levels observed for extracts adjusted to pH 6.0 (about 20% of maximum levels). Tissue extracts supplemented with 9- to 18-fold excess MCSO were subject to modest increases in both extent of discoloration and thiosulfinate levels.

Bioactivities of Kernel Extracts of 18 Strains of Maize (Zea mays)

Aqueous and ethanolic extracts of maize kernels from 18 varieties/strains were prepared for the evaluation of inhibitory activities toward α-glucosidase and scavenging activities toward nitric oxide (NO•) and superoxide (•O(2)(-)). All ethanolic extracts of maize strains tested inhibited yeast (Saccharomyces cerevisiae) α-glucosidase with the highest potency (49% to 54%) found for 2 purple and a yellow strains. However, inhibitory effects of maize extracts on rat intestinal α-glucosidase were as a whole about 10% as effective as with the yeast enzyme. Maize extracts were capable of scavenging NO• at the level of 0.25 mg/mL to extents ranging from 24% to 50% and 26% to 57%, respectively, for aqueous and ethanolic extracts. All tested aqueous extracts were also capable of scavenging •O(2)(-), with efficacies ranging from 8% to 38%, at the level of 1.5 mg/mL, whereas almost none of the ethanolic extracts scavenged •O(2)(-), except for one purple strain (approximately 10% effective). The effectiveness in the enzyme inhibition and antioxidant assays did not correlate with total phenolic and/or anthocyanin levels, nor with the nature of pigmentation among the maize strains evaluated. Practical Application: A diversity of pigmented maize strains was evaluated for biological activities related to mitigating oxidative stress and slowing down glucose absorption from the diet. Certain strains tended to be more abundant in these biological activities and have potential to be used in dietary regimes that are designed to promote human health.