Quantitative Analysis on 2-Acetyl-1-pyrroline of an Aromatic Rice by Stable Isotope Dilution Method and Model Studies on its Formation during Cooking

A quantitative analysis of 2-acetyl-1-pyrroline from aromatic rice samples using a stable isotope dilution method was developed. The compound was extracted from seedlings, roots, and husks at room temperature for 2 h, whereas milled or brown rice samples were extracted at 75 °C. The recovery of 2-acetyl- 1-pyrroline was linear from 5 to 5000 ng/g with sensitivity of less than 0.1 ng/g. 2-Acetyl-1-pyrroline showed tautomerism with animide form. The results revealed that 2-acetyl-1-pyrroline present in aromatic rice samples did not form during cooking or postharvest processes.     

2-Acetyl-1-pyrroline: A key aroma component of aromatic rice and other food products

2-Acetyl-1-pyrroline is an aroma compound that gives aromatic rice its characteristic flavor. This compound either is present naturally in various food sources or is generated during certain processing methods, as reported in a number of studies. This review focuses on several sources of 2-acetyl-1-pyrroline, including aromatic rice, and the factors, including chemical and genetic parameters, affecting the formation of 2-acetyl-1-pyrroline. Extensive work has been conducted on the agricultural parameters, postharvest processing, storage, and cooking methods, influencing the concentrations of 2-acetyl-1-pyrroline in different food commodities. This article is an attempt to emphasize the importance of this compound in the food industry as a major aroma compound.     

Formation of pyrazines and 2-acetyl-1-pyrroline by Bacillus cereus

The production of alkylpyrazines and 2-acetyl-1-pyrroline by different Bacillus cereus strains, which has been previously reported, was studied in detail. B. cereus ATCC 27522 produced the highest amounts of flavour compounds when grown as surface cultures on plate count agar. Pyrazine, methylpyrazine, 2,5-dimethylpyrazine, trimethylpyrazine and 3-ethyl-2,5-dimethylpyrazine were produced in low amounts. Since they were also detected in control flasks, an enzymatic formation was concluded to be unlikely. Only the production of 3-ethyl-2,5-dimethylpyrazine was in all cases significantly different from the control. Detailed precursor studies revealed that the production of 2-acetyl-1-pyrroline by B. cereus ATCC 27522 proceeds via acetylation of 1-pyrroline, a metabolic degradation product of proline and ornithine. Comparison of results obtained from dynamic headspace and simultaneous steam distillation – solvent extraction showed that the use of a non-thermal extraction method is essential to obtain reliable results on the biological formation of these Maillard flavour compounds.     

Quantitative analysis of the isoflavone content and biological growth of soybean (Glycine max L.) at elevated temperature,CO2 level and N application

Effects of ambient and elevated CO₂ levels (360 and 650 µmol mol^?1 respectively), ambient and high (5 °C above ambient) temperatures and their interactions with N application on soybean (Glycine max L.) were studied in 2001. Overall, total isoflavones in whole soybean seeds were highest (1383.0 µg g^?1) in the elevated CO₂ (AE) treatment without N application and lowest (414.1 µg g^?1) in the elevated temperature (EA) treatment with N application. Malonylgenistin (449.2 µg g^?1) and malonyldaidzin (435.9 µg g^?1) concentrations in the AE treatment without N application were highest among the 12 individual isoflavones, while aglycon and acetyl conjugates showed lower concentrations (below 10 µg g^?1) than glucoside and malonyl conjugates in all treatments. Overall, N application had no effect on total isoflavone concentration, while both temperature and CO₂ level had a higher effect on increasing isoflavones, including aglycon and acetyl conjugates (P = 0.001). In the biological growth analysis, total dry weight was highest (100.9 g) in the elevated temperature and CO₂ (EE) treatment with N application, while leaf area was more affected by CO₂ than by temperature and increased with N application. There were larger numbers of pods (99) and seeds (176) per plant in the EE treatment with N application, and generally the AE treatment showed a greater increase in 100‐seed weight (g per 100 seeds) and in pods and seeds per plant than other treatments. Overall, total dry weight was highly affected (P = 0.001) by three main factors, temperature, CO₂ and N application, but the interactions temperature × N and temperature × CO₂× N did not affect total dry weight. Also, total dry weight tended to increase with increasing numbers of pods (r² = 0.93***) and seeds (r² = 0.93***) and larger leaf area (r² = 0.85***). In addition, numbers of pods and seeds were significantly affected (P = 0.01–0.001) by temperature, CO₂ and temperature × CO₂. Generally, elevated CO₂ and temperature did not affect N, P and K concentrations in the seeds but did decrease the concentrations of Ca and Mg, which were increased in the AE treatment. Among the nutrients, Ca and Mg were highly correlated with temperature and CO₂ level. N concentration in the seeds increased with applied N and in particular showed a high increase with elevated temperature and ambient CO₂ (EA treatment). The variation in isoflavones was correlated with temperature (r² = ?0.70**) and CO₂ (r² = 0.67**), while N application was not correlated with isoflavone concentration. Also, Ca (r² = ?0.85***) and Mg (r² = ?0.57*) in the seeds were correlated with variation in isoflavones. This indicated that isoflavones were in higher concentrations under conditions of low temperature and increasing CO₂, which also resulted in low Ca and Mg concentrations in the seeds. The results of this study suggest that the long‐term adaptation of the soybean to growth at elevated CO₂ level and high temperature might potentially increase its isoflavone content, growth and yield. Copyright © 2005 Society of Chemical Industry     

Postharvest changes in physicochemical characteristics and free amino acids content of immature vegetable soya bean (Glycine max L.) grains

The aim of this study was to investigate the changes in physicochemical characteristics and free amino acid profile occurring in immature vegetable soya bbean during postharvest storage under three temperatures [5, 10 and 20 (control), ±1 °C] for 7 days. The results showed that a lower temperature provided an effective control in reducing weight loss, maintained firmness, delayed changes in the pod colour and soluble sugars concentration. Twenty‐two free amino acids in immature soya beans also identified by the ¹H NMR spectroscopy were strongly affected by postharvest temperature, and there were time‐specific differences in the concentration. Those significant differences in free amino acids concentration among storage conditions were closely associated with aspartate and glutamate degradation. Additionally, after 7‐day storage at 5 and 10 °C, the soya bean grains accumulated the highest amount of some essential and flavour amino acids, which revealed storage conditions selected should be dictated by the goal.     

Kinetics of free protein amino acids, free non-protein amino acids and trigonelline in soybean (Glycine max L.) and lupin (Lupinus angustifolius L.) sprouts

A contribution for a better nutritional knowledge of lupin (Lupinus angustifolius L. var. zapaton) and soybean (Glycine max L. var. merit and var. jutro) sprouts about free protein amino acids (FPAA), free non-protein amino acids (FNPAA such as β-N-oxalyl-l-α,β-diaminopropionic acid (β-ODAP), α-aminoadipic acid, γ-aminobutyric acid (GABA) and β-alanine) and trigonelline content has been carried out. Seeds were germinated at 20 °C and darkness during several days in order to obtain good appearance sprouts. The content of FPAA and FNPAA varied among species and varieties. FPAA increased dramatically during germination and the lowest concentrations were found for G. max L. var. jutro. Cys was not detected as FPAA in seeds and seedlings of both legumes. Asn was present in the highest amounts in soybean and lupin seedlings. The α-aminoadipic acid gliotoxin, which was not present in raw soybean seeds, appeared in low amounts during germination. Later stages of germination caused significant increases of β-alanine and GABA, amino acids with beneficial properties. Germination also affected the content of the multifunctional plant hormone trigonelline and slight changes were observed. Highest levels of FPAA, beneficial FNPAA and trigonelline were found at 4, 6 and 9 days of germination for G. max L. var. jutro, G. max L. var. merit and L. angustifolius L. var. zapaton, respectively. Furthermore, these selected germination conditions showed low levels of the gliotoxic α-aminoadipic acid ensuring nutritional safety of the studied seedlings.     

A correlation between the level of phenolic compounds and the antioxidant capacity in cooked-with-rice and vegetable soybean (Glycine max L.) varieties

This study was undertaken to determine the content of phenolic compounds contained in embryo, cotyledon and seed coat of nine soybean (Glycine max L.) varieties. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity was also measured to determine the correlation between the level of phenolic compounds and antioxidant capacity. A total of 10 anthocyanin constituents and 21 phenolic compounds was detected and quantified. In all nine varieties, the seed coat and cotyledon had the highest and lowest levels of phenolic compounds and anthocyanins, respectively despite the fact that all of them showed a wide variation in total amounts of phenolic compounds and anthocyanins in seed coat, embryo, and cotyledon. The seed coat tissue, but not other seed parts, showed a strong correlation between the seed coat color and the content of both phenolic compounds and anthocyanins. The brown and black soybean seed coat contained much higher levels of phenolic compounds and anthocyanins in seed coat tissue than the yellow or green coat soybean. Among the individual phenolic compounds, syringic acid (214 μg g⁻¹) and chlorogenic acid (31 μg g⁻¹) were highest in seed coat and embryo, respectively. Myricetin was highest both in whole seed (16.7 μg g⁻¹) and cotyledon (16.0 μg g⁻¹), being equivalent to 20 and 30% of total phenolic compounds, respectively. Among the 10 anthocyanins, cyanidin-3-glucoside was found to accumulate at the highest level in the seed coat (1783 μg g⁻¹), whole seed (106 μg g⁻¹) and embryo (0.35 μg g⁻¹), which correspond to 95, 96, and 40% of the total anthocyanin contents, respectively. The cotyledon accumulated pelargonidin-3-glucoside (0.39 μg g⁻¹) at the highest level that is equivalent to 62% of total anthocyanin contents. DPPH activity was found to have a strong correlation (^***probability < 0.001) with phenolic compounds (0.67^***) and anthocyanins (0.70^***).J. A. Kim, W. S. Jung, and S. C. Chun are equally contributed to this work.     

Influence of plant growth regulators on key-coding genes expression associated with phytochemicals biosynthesis and antioxidant activity in soybean (Glycine max (L.) Merr) sprouts

The present study was aimed to investigate the effect of plant growth regulators (PGRs) on phytochemicals biosynthesis and antioxidant activity in soybean sprouts. _GmVTC2, _GmDHAR and _GmGLDH were investigated for the biosynthesis of vitamin C. _GmPAL, _GmCHS and _GmIFS were investigated for the biosynthesis of phenolics, flavonoids and isoflavones. Our results revealed that 6-benzylaminopurine (6-BA) treatment significantly affected the expression levels of those major genes, followed by Gibberellin (GA3) and Naphthaleneacetic acid (NAA). Moreover, the highest levels of vitamin C, polyphenolics and antioxidant activity in 6-BA and GA3 treated groups indicate the synergistic and antagonistic role of growth regulators in the biosynthesis of phytochemicals. Overall, 6-BA seems the key contributor in genes’ expression involve in the biosynthesis secondary metabolites and their antioxidant activity. Therefore, it could be the most suitable option to enhance nutritional quantity and health benefits of soybean sprouts.     

Comparison of post‐germination mobilization of cell wall polysaccharides and non‐cell wall carbohydrates in soybean (Glycine max L.) cotyledons

BACKGROUND: In previous research, we demonstrated that cell wall polysaccharide (CWP) levels of soybean (Glycine max L.) cotyledons are negatively correlated with the sum of seed oil and protein content. Although the results suggest that reducing cotyledon CWP levels would be desirable, it is not known whether CWP are mobilized during early seedling growth and, if so, to what extent mobilization contributes to seed reserves. RESULTS: Ungerminated (dry) seeds contained equivalent amounts [approximately 20 mg (cotyledon pair)^?1] of non‐cell wall carbohydrates (NCWC) and CWP. Galactose and arabinose accounted for 47% of total CWP in cotyledons of dry seeds. Measured 14 days after planting (DAP), the levels of NCWC and CWP were reduced 98% and 34%, respectively, in cotyledons of seedlings grown under a 16‐h photoperiod. Measured 14 DAP, greater than 85% of cotyledon cell wall galactose plus arabinose was mobilized. The transformation of the cotyledon to a photosynthetic organ was associated with restructuring of the cell wall involving increases in uronic acids, glucose and rhamnose. CONCLUSION: CWP of soybean cotyledons are modified during early seedling growth due to mobilization and cell wall restructuring triggered by light. The amount of carbon mobilized makes only a small contribution to total cotyledon reserves. Published 2009 by John Wiley & Sons, Ltd.     

Antinutritional and/or toxic factors in soybean (Glycine max (L) Merril) seeds: comparison of different cultivars adapted to the southern region of Brazil

Soybean (Glycine max (L) Merril) seeds are known to contain different proteins displaying antinutritional and/or toxic effects, such as soybean agglutinin (an N‐acetylgalactosamine‐specific lectin), proteinase inhibitors (Kunitz‐ and Bowman–Birk‐type trypsin and chymotrypsin inhibitors) and urease (seed and tissue isoforms). Two other toxic proteins were previously isolated from soybeans, soyatoxin (21 kDa) and soybean toxin (18.4 kDa), which are immunologically related to canatoxin, a toxic protein from Canavalia ensiformis (jackbean) seeds. In this work we have screened crude extracts from seeds of six different soybean cultivars, which together represent most of the crop harvested in the southern region of Brazil, for the presence of urease, trypsin inhibitory and haemagglutination activities, intraperitoneal toxicity in mice and immunoreactivity against anti‐canatoxin antibodies. Significant differences were found in the contents of proteinase inhibitors, lectin, urease activity and lethality in mice. The relevance of these findings to the agronomic qualities and to the choice of soybean cultivars to be used as food or feed is discussed. Copyright © 2004 Society of Chemical Industry     

Thirty‐three years of 2‐acetyl‐1‐pyrroline,a principal basmati aroma compound in scented rice (Oryza sativa L.): a status review

Rice is the staple food of around 3 billion people, most of them in Asia which accounts for 90% of global rice consumption. Aromatic rices have been preferred over non‐aromatic rice for hundreds of years. They have a premium value in national as well as international market owing to their unique aroma and quality. Many researchers were involved in identifying the compound responsible for the pleasant aroma in aromatic rice in the 20th century. However, due to its unstable nature, 2‐acetyl‐1‐pyrroline (2AP) was discovered very late, in 1982. Buttery and co‐workers found 2AP to be the principal compound imparting the pleasant aroma to basmati and other scented rice varieties. Since then, 2AP has been identified in all fragrant rice (Oryza sativa L.) varieties and a wide range of plants, animals, fungi, bacteria and various food products. The present article reviews in detail biochemical and genetic aspects of 2AP in living systems. The site of synthesis, site of storage and stability in plant systems in vivo is of interest. This compound requires more research on stability to facilitate use as a food additive. © 2016 Society of Chemical Industry     

Hydrolysis of the toxic constituents (vicine and convicine) in fababean (Vicia faba L.) food preparations following treatment with β-glucosidase

Several experiments were carried out to determine whether the concentrations of vicine and convicine in fababean food preparations were reduced following heat and β-glucosidase treatment. Heating fababeans to relatively high temperatures was not effective in reducing the concentration of these glycosides. Treatment of cooked fababeans (fababean paste) with ground raw almond powder, which is a rich source of β-glucosidase (EC 3.2.1.21) activity, resulted in the hydrolysis of vicine and convicine. The degree of hydrolysis ranged from partial to complete and was dependent on the duration of the treatment period (1 to 24 h), the temperature at which treatment was carried out (1 to 30°C), the pH (6.4 to 4.6) as adjusted by the addition of lemon juice and the amount of ground almonds added to the fababean paste. For example, 88 to 89% of the glycosides were hydrolysed when 1 g of fababean paste was mixed with 0.1 g of almond powder and 0.1 ml of lemon juice and incubated at 30°C for 3 h. This study demonstrated that the concentration of vicine and convicine and their corresponding aglycones can be greatly reduced in fababean food preparations by enzymic hydrolysis.     

Polygonum viviparum L. inhibits the lipopolysaccharide‐induced inflammatory response in RAW264.7 macrophages through haem oxygenase‐1 induction and activation of the Nrf2 pathway

BACKGROUND: Polygonum viviparum L. (PV) is a member of the family Polygonaceae and is widely distributed in high‐elevation areas. It is used as a folk remedy to treat inflammation‐related diseases. This study was focused on the anti‐inflammatory response of PV against lipopolysaccharide (LPS)‐induced inflammation in RAW264.7 macrophages. RESULTS: Treatment with PV did not cause cytotoxicity at 0–50 µg mL^?1 in RAW264.7 macrophages, and the IC₅₀ value was 270 µg mL^?1. PV inhibited LPS‐stimulated nitric oxide (NO), prostaglandin (PG)E₂, interleukin (IL)‐1β and tumour necrosis factor (TNF)‐α release and inducible NO synthase (iNOS) and cyclooxygenase (COX)‐2 protein expression. In addition, PV suppressed the LPS‐induced p65 expression of nuclear factor (NF)‐κB, which is associated with the inhibition of IκB‐α degradation. These results suggest that, among mechanisms of the anti‐inflammatory response, PV inhibits the production of NO and these cytokines by down‐regulating iNOS and COX‐2 gene expression. Furthermore, PV can induce haem oxygenase (HO)‐1 protein expression through nuclear factor E2‐related factor 2 (Nrf2) activation. A specific inhibitor of HO‐1, zinc(II) protoporphyrin IX, inhibited the suppression of iNOS and COX‐2 expression by PV. CONCLUSION: These results suggest that PV possesses anti‐inflammatory actions in macrophages and works through a novel mechanism involving Nrf2 actions and HO‐1. Thus PV could be considered for application as a potential therapeutic approach for inflammation‐associated disorders. © 2012 Society of Chemical Industry     

Comparative formation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in creatinine/phenylalanine and creatinine/phenylalanine/4-oxo-2-nonenal reaction mixtures

The comparative formation of the heterocyclic aromatic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in both creatinine/phenylalanine (CRN/Phe) and creatinine/phenylalanine/4-oxo-2-nonenal (CRN/Phe/ON) systems was studied to analyse the ability of lipid-derived reactive carbonyls to promote PhIP formation. Although PhIP was produced to some extent in the CRN/Phe system, the presence of the oxidized lipid increased considerably the amount of PhIP produced. This increase seemed to be a consequence of the decrease in the E_a of the reaction when the lipid was present, which diminished from 112.9 to 80.9 kJ/mol. On the other hand, the addition of the lipid did not seem to produce PhIP by an alternative mechanism because PhIP was formed analogously in both CRN/Phe and CRN/Phe/ON systems as a function of pH, creatinine concentration, phenylalanine concentration, time, temperature, oxygen concentration in the reaction atmosphere, and the addition of different amounts of ammonia. All these results suggest that the ability of lipid oxidation products to produce PhIP is related to their capacity to induce the Strecker degradation of phenylalanine to phenylacetaldehyde. Therefore, any other reactive carbonyl compound that can produce the Strecker degradation of phenylalanine should also be considered as a potential inducer of PhIP formation under appropriate conditions.     

Site-specific mutagenesis of the N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine DNA adduct in mammalian cells

 Site-specifically modified oligodeoxynucleotides were used to explore the mutagenic properties of a cooked food mutagen-derived DNA adduct, N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-PhIP). A dG-C8-PhIP-modified oligodeoxynucleotide was prepared post-synthetically by reacting an oligodeoxynucleotide containing a single dG (^5′-TCC TCC TCG CCT CTC T) with N-acetoxy-PhIP. The unmodified and dG-C8-PhIP-modified oligomers were inserted into single-strand (ss) phagemid vectors. These ss vectors were transfected into simian kidney (COS-7) cells. The progeny plasmid obtained was used to transform Escherichia coli DH10B. The transformants were analyzed by oligodeoxynucleotide hybridization and sequencing to determine the mutation frequency and spectrum. Preferential incorporation of dCMP, the correct base, was observed opposite dG-C8-PhIP. Targeted mutants showing G→T transversions were detected, along with a small number of G→A transitions and G→C transversions. Significant amounts of non-targeted mutations representing C→T transitions also were detected 5′ to the dG-C8-PhIP lesion. Thus, dG-C8-PhIP, a major DNA adduct induced by PhIP, is mutagenic in mammalian cells and may be involved in the initiation of human cancer. Received: 23 April 1998     

Site-specific mutagenesis of the N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine DNA adduct in mammalian cells

 Site-specifically modified oligodeoxynucleotides were used to explore the mutagenic properties of a cooked food mutagen-derived DNA adduct, N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-PhIP). A dG-C8-PhIP-modified oligodeoxynucleotide was prepared post-synthetically by reacting an oligodeoxynucleotide containing a single dG (^5′-TCC TCC TCG CCT CTC T) with N-acetoxy-PhIP. The unmodified and dG-C8-PhIP-modified oligomers were inserted into single-strand (ss) phagemid vectors. These ss vectors were transfected into simian kidney (COS-7) cells. The progeny plasmid obtained was used to transform Escherichia coli DH10B. The transformants were analyzed by oligodeoxynucleotide hybridization and sequencing to determine the mutation frequency and spectrum. Preferential incorporation of dCMP, the correct base, was observed opposite dG-C8-PhIP. Targeted mutants showing G→T transversions were detected, along with a small number of G→A transitions and G→C transversions. Significant amounts of non-targeted mutations representing C→T transitions also were detected 5′ to the dG-C8-PhIP lesion. Thus, dG-C8-PhIP, a major DNA adduct induced by PhIP, is mutagenic in mammalian cells and may be involved in the initiation of human cancer. Received: 23 April 1998