Reaction of tryptophan with carbohydrates: mechanistic studies on the formation of carbohydrate-derived beta-carbolines.

Numerous carbohydrate-derived beta-carbolines have been identified for the first time in model reactions of tryptophan with glucose and ribose, as well as in food samples. Extending these structural studies, we performed detailed investigations to elucidate the corresponding intermediates and formation pathway of these alkaloids. Degradation experiments with purified tryptophan glycoconjugates established that only glyco-tetrahydro-beta-carboline-3-carboxylic acids, and not the N-glycosides nor the C-glycoconjugates represented the direct precursors of carbohydrate-derived beta-carbolines. In addition, the significance of the oxidative decarboxylation reaction as the initial step for formation of 1-substituted beta-carbolines was proven. Finally, the stereochemistry of the carbohydrate-derived side chain was studied by means of CD spectroscopy and HPLC-CD experiments. These detailed stereochemical analyses yielded experimental evidence for the racemization steps required for formation of the carbohydrate-derived harman alkaloids and confirmed the proposed reaction pathway.     

Enzymatic formation of ether linkage producing shoyuflavones from genistein and (+/-)-trans-epoxysuccinic acid

The production mechanism of shoyuflavones, conjugated ethers of isoflavones with tartaric acid and isolated from fermented soy sauce, was studied. In the high molecular weight fraction of the culture extract of Aspergillus oryzae, genistein was transformed into shoyuflavone B in the presence of (+/-)-trans-epoxysuccinic acid but not in the low molecular one. Asp. sojae and Asp. tamarii showed high activity similar to Asp. oryzae but none of Asp. niger, Rhizopus oligosporus, and Mucor praini did. The contents of epoxysuccinic acids in the starting materials of soy sauce and the cultures of various Asp. fungi were determined as dimethyl 2-chloro-3-hydroxysuccinate derivatives by GC-MS. Although epoxysuccinic acids were contained in Asp. oryzae, Asp. sojae, and Asp. tamarii cultures, they were not found in soybeans and wheat. A possible producing mechanism for shoyuflavones by enzymatically conjugating isoflavones to (+/-)-trans-epoxysuccinic acid with ether linkage was suggested.     

Biodegradation of oxalic acid from spinach using cereal radicles

A high level of oxalate intake constitutes a health risk for infants and metabolically disposed adults. Spinach, acclaimed for its many health benefits, is among the vegetables richest in oxalate. Blanching reduces oxalate unsatisfactorily and unspecifically. An alternative, biological method is proposed on the basis of rye seedlings or radicles (also barley and wheat) containing an oxalate-specific oxalate oxidase by nature. Dissolved oxalate (0.25 mM) was rapidly degraded in the presence of radicles (e.g., 70% within 100 min). With commercial deep-frozen spinach, near-complete degradation of soluble oxalate was achieved at pH 3.5. The total level of oxalate was reduced by half. Similarly high rates occurred from 18 to 35 degrees C. Even at 55 degrees C appreciable rates were observed. The seedling as a whole is effective, too, and enrichment with cereal-specific healthy components would occur. Removal of oxalate from other vegetables, juices, cycled process waters, or feeds is conceivable with fresh or heat-dried cereal seedlings or radicles.     

菠菜叶片硝态氮还原对叶柄硝态氮含量的影响

选用3个菠菜品种,设置N.0.1和0.3.g/kg2个施氮水平进行盆栽试验。在不同时期采样测定叶片内、外源硝酸还原酶活性、硝态氮代谢/贮存库大小,以及加入外源硝态氮培养后叶片硝酸还原酶活性的变化,探讨菠菜叶片的硝态氮还原与叶柄硝态氮含量的关系。结果表明,叶片内源硝酸还原酶活性、内源/外源硝酸还原酶活性比值、叶片的硝态氮代谢库大小及代谢/贮存库比值与叶柄硝态氮含量呈相反趋势。加入外源硝态氮培养后叶片硝酸还原酶活性的增加程度与叶柄硝态氮含量相一致。叶片内源硝酸还原酶活性高低及其发挥程度,叶片硝态氮代谢库大小及硝态氮在代谢、贮存库中的分配是造成品种间叶柄硝态氮含量高低差异的重要原因。     

黄腐酸和聚天冬氨酸对蕹菜氮素吸收及氮肥去向的影响

为了解黄腐酸(FA)和聚天冬氨酸(PASP)对蕹菜氮素吸收及氮肥去向的影响,采用15N尿素示踪技术,设置不施氮肥(CK),单施尿素(N),尿素配施低、中、高用量的FA和PASP(NF1、NF2、NF3、NP1、NP2、NP3)8个处理,在温室条件下进行盆栽试验。结果表明,与N处理相比,配施FA和PASP后蕹菜地上部鲜重增加了7.46%~17.55%;NP2、NP3和各NF处理显著提高了蕹菜的吸氮量,提高幅度为10.84%~18.25%,其中,蕹菜对非标记氮的吸收量显著增加,且随FA和PASP用量的增大而增加;NF3处理的15N利用率显著低于N处理,其余处理无显著变化;NF2、NF3、NP2和NP3处理的15N损失率比N处理减少了5.41~14.58个百分点;NF2、NF3和NP2处理的15N土壤残留率增加了5.08~20.02个百分点。研究表明,中、高用量的FA和PASP与尿素配施促进了蕹菜对氮素(尤其是非标记氮素)的吸收,同时减少了氮肥的损失,增加了氮肥在土壤中的残留,对土壤氮库的贡献作用显著。     

Origin and mechanistic pathways of formation of the parent furan--a food toxicant

Studies performed on model systems using pyrolysis-GC-MS analysis and (13)C-labeled sugars and amino acids in addition to ascorbic acid have indicated that certain amino acids such as serine and cysteine can degrade and produce acetaldehyde and glycolaldehyde that can undergo aldol condensation to produce furan after cyclization and dehydration steps. Other amino acids such as aspartic acid, threonine, and alpha-alanine can degrade and produce only acetaldehyde and thus need sugars as a source of glycolaldehyde to generate furan. On the other hand, monosaccharides are also known to undergo degradation to produce both acetaldehyde and glycolaldehyde; however, (13)C-labeling studies have revealed that hexoses in general will mainly degrade into the following aldotetrose derivatives to produce the parent furan-aldotetrose itself, incorporating the C3-C4-C5-C6 carbon chain of glucose (70%); 2-deoxy-3-ketoaldotetrose; incorporating the C1-C2-C3-C4 carbon chain of glucose (15%); and 2-deoxyaldotetrose, incorporating the C2-C3-C4-C5 carbon chain of glucose (15%). Furthermore, it was also proposed that under nonoxidative conditions of pyrolysis, ascorbic acid can generate the 2-deoxyaldotetrose moiety, a direct precursor of the parent furan. In addition, 4-hydroxy-2-butenal-a known decomposition product of lipid peroxidation-was proposed as a precursor of furan originating from polyunsaturated fatty acids. Among the model systems studied, ascorbic acid had the highest potential to produce furan, followed by glycolaldehyde/alanine > erythrose > ribose/serine > sucrose/serine > fructose/serine > glucose/cysteine.     

应用超弱光子辐射评价菠菜叶片衰老方法可行性

为了研究植物叶片衰老评价及其无损检测方法,采用外源H2O2诱发菠菜叶片衰老,研究了H2O2胁迫下菠菜叶片细胞超弱光子辐射的变化规律及其与叶片衰老的关系。结果表明,在H2O2胁迫过程中,菠菜叶片自发光子辐射随着胁迫时间的延长而增加,外界光诱导的延迟光子辐射也表现出明显的变化。根据细胞自发光子辐射和延迟光子辐射的生物学意义,采用延迟光子辐射积分强度和自发光子辐射的比值作为描述细胞整体状态的状态参量,结果发现,在H2O2胁迫过程中,基于细胞超弱光子辐射的菠菜叶片细胞状态参量的变化与叶绿素含量的变化呈现良好的正相关,相关系数为0.91437,暗示通过细胞状态参量的变化可以推测菠菜叶片细胞的状态与衰老程度,进而实现对菠菜叶片衰老程度的无损检测,为植物叶片衰老评价提供了一种新方法。     

Antioxidant capacity and phenolic content in leaf extracts of tree spinach (Cnidoscolus spp.)

Total phenolic content and antioxidant capacity of two tree spinach species (Cnidoscolus chayamansa McVaugh and C. aconitifolius Miller.) were determined in raw and cooked leaf extracts. Antioxidant capacity was assessed by the oxygen radical absorbance capacity (ORAC) assay, and flavonoid glycoside composition was quantified by HPLC and identified by GC. Total phenolics and antioxidant capacity were higher in raw than in cooked leaf extracts. The ORAC values were strongly correlated with total phenolic content (r = 0.926) in all leaf extracts. The major flavonoids isolated from the leaf extracts were kaempferol-3-O-glycosides and quercetin-3-O-glycosides. C. aconitifolius leaves contained more varieties of the flavonoid glycosides than C. chayamansa. Cooking reduced antioxidant activity and phenolic content and resulted in losses of some kaempferol glycoside and quercetin glycoside residues in leaf extracts. The results of this study indicate that tree spinach leaves are a rich source of natural antioxidants for foods.     

In-depth mechanistic study on the formation of acrylamide and other vinylogous compounds by the maillard reaction

The formation of acrylamide was studied in low-moisture Maillard model systems (180 degrees C, 5 min) based on asparagine, reducing sugars, Maillard intermediates, and sugar degradation products. We show evidence that certain glycoconjugates play a major role in acrylamide formation. The N-glycosyl of asparagine generated about 2.4 mmol/mol acrylamide, compared to 0.1-0.2 mmol/mol obtained with alpha-dicarbonyls and the Amadori compound of asparagine. 3-Hydroxypropanamide, the Strecker alcohol of asparagine, generated only low amounts of acrylamide ( approximately 0.23 mmol/mol), while hydroxyacetone increased the acrylamide yields to more than 4 mmol/mol, indicating that alpha-hydroxy carbonyls are much more efficient than alpha-dicarbonyls in converting asparagine into acrylamide. The experimental results are consistent with the reaction mechanism based on (i) a Strecker type degradation of the Schiff base leading to azomethine ylides, followed by (ii) a beta-elimination reaction of the decarboxylated Amadori compound to afford acrylamide. The beta-position on both sides of the nitrogen atom is crucial. Rearrangement of the azomethine ylide to the decarboxylated Amadori compound is the key step, which is favored if the carbonyl moiety contains a hydroxyl group in beta-position to the nitrogen atom. The beta-elimination step in the amino acid moiety was demonstrated by reacting under low moisture conditions decarboxylated model Amadori compounds obtained by synthesis. The corresponding vinylogous compounds were only generated if a beta-proton was available, for example, styrene from the decarboxylated Amadori compound of phenylalanine. Therefore, it is suggested that this thermal pathway may be common to other amino acids, resulting under certain conditions in their respective vinylogous reaction products.     

Model studies on the oxygen-induced formation of benzaldehyde from phenylacetaldehyde using pyrolysis GC-MS and FTIR

Benzaldehyde, a potent aroma chemical of bitter almond, can also be formed thermally from phenylalanine and may contribute to the formation of off-aroma. To identify the precursors involved in its generation during Maillard reaction, various model systems containing phenylalanine, phenylpyruvic acid, phenethylamine, or phenylacetaldehyde were studied in the presence and absence of moisture using oxidative and nonoxidative Py-GC-MS. Analysis of the data indicated that phenylacetaldehyde, the Strecker aldehyde of phenylalanine, is the most effective precursor and that both air and water significantly enhanced the rate of benzaldehyde formation from phenylacetaldehyde. Phenylpyruvic acid was the most efficient precursor under nonoxidative conditions. Phenethylamine, on the other hand, needed the presence of a carbonyl compound to generate benzaldehyde only under oxidative conditions. On the basis of the results obtained, a free radical initiated oxidative cleavage of the carbon-carbon double bond of the enolized phenylacetaldehyde was proposed as a possible major mechanism for benzaldehyde formation, and supporting evidence was provided through monitoring of the evolution of the benzaldehyde band from heated phenylacetaldehyde in the presence and absence of 1,1'-azobis(cyclohexanecarbonitrile) on the ATR crystal of an FTIR spectrophotometer. In the presence of the free radical initiator, the enol band of the phenylacetaldehyde centered at 1684 cm(-1) formed and increased over time, and after 18 min of heating time the benzaldehyde band centered at 1697 cm(-1) formed and increased at the expense of the enol band of phenylacetaldehyde, indicating a precursor product relationship.     

Impact of the N-terminal amino acid on the formation of pyrazines from peptides in Maillard model systems

Only a minor part of Maillard reaction studies in the literature focused on the reaction between carbohydrates and peptides. Therefore, in continuation of a previous study in which the influence of the peptide C-terminal amino acid was investigated, this study focused on the influence of the peptide N-terminal amino acid on the production of pyrazines in model reactions of glucose, methylglyoxal, or glyoxal. Nine different dipeptides and three tripeptides were selected. It was shown that the structure of the N-terminal amino acid is determinative for the overall pyrazine production. Especially, the production of 2,5(6)-dimethylpyrazine and trimethylpyrazine was low in the case of proline, valine, or leucine at the N-terminus, whereas it was very high for glycine, alanine, or serine. In contrast to the alkyl-substituted pyrazines, unsubstituted pyrazine was always produced more in the case of experiments with free amino acids. It is clear that different mechanisms must be responsible for this observation. This study clearly illustrates the capability of peptides to produce flavor compounds such as pyrazines.     

Biosynthesis of menthofuran in Mentha x piperita: stereoselective and mechanistic studies.

Mentha x piperita shoot tips and first leaf pair were fed with aqueous solutions of [(2)H(2)]- and [(2)H(2)]/[(18)O]-labeled pulegone. The essential oil was analyzed by solid phase microextraction and enantioselective multidimensional gas chromatography/mass spectrometry. After feeding experiments with labeled pulegone racemate, both labeled (S)-menthofuran and (R)-menthofuran were detectable simultaneously together with genuine (R)-menthofuran. It could be shown that both labeled pulegone enantiomers are converted by Mentha x piperita to the corresponding labeled menthofuran enantiomers, favoring the labeled analogue of the nongenuine (S)-pulegone. The oxygen in menthofuran is introduced by enzymatic oxidation of pulegone, as concluded from feeding experiments with mixed labeled [(2)H(2)]/[(18)O]pulegone.     

硼砂叶面喷施处理对马铃薯产量形成的影响

以"克新13"马铃薯为试材,采用不同浓度的硼砂叶面喷施,通过测定马铃薯地上部干重、地下部干重、根系活力、叶绿素含量及产量等指标,研究硼砂对马铃薯产量形成的作用效应。结果表明:适量的硼砂叶面喷施能有效地提高根系活力、叶绿素含量,促进马铃薯地上部和地下部干物质积累,从而促进产量的形成。     

Model studies on the formation of monochloropropanediols in the presence of lipase

The formation of chloropropanols was investigated using model systems comprised of lipase, vegetable oil or fat, water, and sodium chloride. The results showed that measurable levels of the foodborne carcinogen 3-chloro-1,2-propanediol (3-MCPD) are formed in the presence of commercially available lipases of mammalian, vegetable, and fungal origins, incubated at temperatures of 40 degrees C. The highest yield of 3-MCPD was obtained in reaction mixtures containing lipase from Rhizopus oryzae, and all the lipases studied exhibited a high hydrolytic activity toward triglycerides from palm and peanut oil. In contrast, hydrolysis over time and the yield of 3-MCPD in olive and sunflower oils were significantly lower (up to 10-fold), possibly linked to the relatively lower amount (<18%) of saturated fatty acids in these oils. We provide here for the first time evidence that lipases are able to induce the formation of chloropropanols under model system conditions. However, the key intermediates and precise mechanistic aspects governing the formation of 3-MCPD in the presence of lipase still need to be elucidated.     

低次烟叶中蛋白质提取工艺优化及氨基酸分析研究

为了充分综合利用低次烟叶蛋白资源,采用碱溶酸沉法对低次烟叶中蛋白质的提取工艺进行了研究,并利用高效液相色谱系统对其氨基酸组成进行了分析。研究结果表明：烟叶磨浆的最佳工艺为固液比1∶17,提取温度60℃,提取液pH值8.0,磨浆两次;烟叶蛋白碱溶最佳工艺为温度60℃,pH8.0,时间60 min,搅拌条件下碱溶提取3次;烟叶蛋白酸沉工艺中,在pH3.0,4℃下静置8 h效果最佳,烟叶蛋白酸沉提取率最高为86.71％;烟叶蛋白的氨基酸分析结果显示,烟叶蛋白的氨基酸组成种类齐全,必需氨基酸含量较高,氨基酸分数较高,表明低次烟叶蛋白是一类比较优质的蛋白质资源。     

Environmental effects on the elemental leaf composition of crassulacean acid metabolism plants

Environmental effects on several CAM plants which bring them in CAM (when plants show diurnal fluctuations in the concn of titratable acid) and out of CAM (when acid concn remain low with no diurnal fluctuation) were studied. Change in photoperiod alone caused the greatest amplitude of the acid fluctuation from 2.6 to 2.7 meq acid/100 g FW in Sedum telephoides. Light intensity affected the K/Ca ratio from a high when in CAM to a low when out of CAM in the 2 species. The plasma emission spectrograph analysis of the leaf tissue grown in the specific environment showed the presence of high levels of Mn, B, Cu, Zn, Na, Al and Si in Sedum telephoides. The elemental analyses of CAM plants showed 1.8 to 9.4% K.     

壳聚糖没食子酸衍生物酶法制备及对鲜切苹果的保鲜效果

为提高壳聚糖水溶性和保鲜性能,该文以没食子酸为配体,漆酶为催化剂催化壳聚糖与没食子酸发生接枝反应,得到壳聚糖没食子酸衍生物,对其结构进行了初步表征,并初步探讨了其对鲜切苹果的保鲜效果。结果表明,衍生物接枝率为72.80%,溶解度达到92.77%,没食子酸中的羧基与壳聚糖分子的氨基发生了迈克尔加成反应,在壳聚糖的C2位生成了酰胺键。14℃下贮藏4 d后,10 mg/m L衍生物处理的苹果硬度损失率仅为10.23%,比空白组、壳聚糖处理组和壳聚糖没食子酸物理复合物处理组分别低20.90%(P0.05)、16.60%(P0.05)和15.60%(P0.05);可溶性固形物、维生素C、谷胱甘肽和多酚质量分数分别为12.93%,1.23 mg/(100 g),6.26 mg/(100 g)和6.24 mg/(100 g),分别比空白组高16.47%(P0.05),48.78%(P0.05),24.92%(P0.05)和43.75%(P0.05);多酚氧化酶和过氧化物酶活性分别比空白组低36.73%(P0.05)和76.94%(P0.05);菌落总数为16.00×10~4 CFU/g,显著低于空白组、壳聚糖处理组和壳聚糖没食子酸物理复合物处理组(P0.05)。结果表明,漆酶可催化壳聚糖与没食子酸发生接枝反应,得到的壳聚糖没食子酸衍生物对鲜切苹果具有较好的保鲜效果。     

Sugar fragmentation in the maillard reaction cascade: isotope labeling studies on the formation of acetic acid by a hydrolytic beta-dicarbonyl cleavage mechanism

The formation of acetic acid was elucidated based on volatile reaction products and related nonvolatile key intermediates. The origin and yield of acetic acid were determined under well-controlled conditions (90-120 degrees C, pH 6-8). Experiments with various 13C-labeled glucose isotopomers in the presence of glycine revealed all six carbon atoms being incorporated into acetic acid: C-1/C-2 ( approximately 70%), C-3/C-4 ( approximately 10%), and C-5/C-6 (approximately 20%). Acetic acid is a good marker of the 2,3-enolization pathway since it is almost exclusively formed from 1-deoxy-2,3-diulose intermediates. Depending on the pH, the acetic acid conversion yield reached 85 mol % when using 1-deoxy-2,3-hexodiulose (1) as a precursor. Hydrolytic beta-dicarbonyl cleavage of 1-deoxy-2,4-hexodiuloses was shown to be the major pathway leading to acetic acid from glucose without the intermediacy of any oxidizing agents. The presence of key intermediates was corroborated for the first time, i.e., tetroses and 2-hydroxy-3-oxobutanal, a tautomer of 1-hydroxy-2,3-butanedione, also referred to as 1-deoxy-2,3-tetrodiulose. The hydrolytic beta-dicarbonyl cleavage represents a general pathway to organic acids, which corresponds to an acyloin cleavage or a retro-Claisen type reaction. Although alternative mechanisms must exist, the frequently reported hydrolytic alpha-dicarbonyl cleavage of 1 can be ruled out as a pathway forming carboxylic acids.     

Tyrosinase-aided protein cross-linking: effects on gel formation of chicken breast myofibrils and texture and water-holding of chicken breast meat homogenate gels

The effects of Trichoderma reesei tyrosinase-catalyzed cross-linking of isolated chicken breast myofibril proteins as a simplified model system were studied with special emphasis on the thermal stability and gel formation of myofibrillar proteins. In addition, tyrosinase-catalyzed cross-linking was utilized to modify the firmness, water-holding capacity (WHC), and microstructure of cooked chicken breast meat homogenate gels. According to SDS-PAGE, the myosin heavy chain (MHC) and troponin T were the most sensitive proteins to the action of tyrosinase, whereas actin was not affected to the same extent. Calorimetric enthalpy (DeltaH) of the major thermal transition associated with myosin denaturation was reduced and with actin denaturation increased in the presence of tyrosinase. Low-amplitude viscoelastic measurements at constant temperatures of 25 degrees C and 40 degrees C showed that tyrosinase substantially increased the storage modulus (G') of the 4% myofibrillar protein suspension in the 0.35 M NaCl concentration. The effect was the most pronounced with high-enzyme dosages and at 40 degrees C. Without tyrosinase, the G' increase was low. Tyrosinase increased the firmness of the cooked phosphate-free and low-meat chicken breast meat homogenate gels compared to the corresponding controls. Tyrosinase maintained gel firmness at the control level of the low-salt homogenate gel and weakened it when both salt and phosphate levels were low. Tyrosinase improved the WHC of the low-meat and low-salt homogenate gels and maintained it at the level of the corresponding controls of phosphate-free and low-salt/low-phosphate homogenate gels. Microstructural characterization showed that a collagen network was formed in the presence of tyrosinase. Keywords: Chicken myofibrillar proteins; protein modification; cross-linking; tyrosinase; gelation; thermal stability; texture; water-holding capacity; microstructure.