Skip to main content
PMC home page
Journal of Food Science and Technology logoLink to Journal of Food Science and Technology
. 2016 Dec 3;53(12):4141–4148. doi: 10.1007/s13197-016-2384-8

Amino acids profile and antioxidant activity of different Lupinus angustifolius seeds after solid state and submerged fermentations

Vytaute Starkute 1, Elena Bartkiene 1,✉, Vadims Bartkevics 2,3, Janis Rusko 3, Daiva Zadeike 4, Grazina Juodeikiene 4
PMCID: PMC5223248  PMID: 28115754

Abstract

The objective of this study was to investigate the amino acids profile, total phenolic compounds (TPC) content, antioxidant activity after submerged (SmF) and solid state (SSF) fermentations of different Lupinus angustifolius seeds by the Lactobacillus sakei KTU05-6. Additionally, the impact of different lupin seeds as fermentation media for LAB biomass and d/l-lactic acid production was analysed. The d/l ratio for SmF and SSF treated lupin samples varied from 0.15 to 0.45 and from 0.12 to 0.46, 16 respectively. Nutritional analysis highlighted a substantial increase in the TPC content and antioxidant activity up to 31.5–48.8% for SSF treated L. angustifolius samples compared to unfermented. The interaction between analysed factors (lupin variety and fermentation conditions) had a significant influence on essential and nonessential amino acids profile.

Keywords: Lupinus angustifolius, Lactobacillus sakei, Amino acids, Phenolic compounds, Antioxidant activity

Introduction

In most parts of the world lupin has traditionally been used primarily as feedstock, and an interest in unique nutritional values of lupin components as food ingredients has increased as people have become more aware of their health benefits. The lupin have a high content of essential amino acids, protein, fibre and low fat content compared to soya bean (Bähr et al. 2015; Thambiraj et al. 2015), that provide the huge industrial potential for this legume. The consumption of lupin may have cholesterol lowering effect and has high antioxidant properties. The potential health benefits of lupin nutritional components increased the potential of lupin incorporation as additive in various food products including pasta (Jayasena and Nasar-Abbas 2012), bakery products (Abdelrahman 2014), wheat bread (Villarino et al. 2015), muffins (Rumiyati et al. 2015).

Legume crops, which represent the major food and feed sources worldwide, contains limiting levels of some essential amino acids (EAA), particularly lysine and methionine (Galili and Amir 2013). The EAA could be complemented from the animal food (meat, eggs and milk) as well as from a variety of cereals and legumes that provide optimal levels of EAA (Galili 2011). Amino acids such as lysine, methionine, threonine, phenylalanine, tryptophan, valine, isoleucine, leucine, and histidine are covered in this definition (Galili et al. 2016). Lupin is considered as a cheap alternative stock to other legume crops such as soya beans as it contains comparable quantity of proteins with similar AA profile.

Legume proteins are resistant to proteolysis because of specific structural properties. Their structural stability affects in vivo digestibility and availability of the EAA, and also the production of bioactive compounds with functional properties (Orlovskaya et al. 2010). The possibility to modulate protein structure during technological processing may be used as a novel strategy to improve the nutritional potential of protein rich plant foods (Carbonaro et al. 2015).

In addition to their favourable nutritional profile, lupin seeds contain significant amounts of polyphenols, carotenoids, phytosterols, tocopherols and peptides with antioxidant, antimicrobial, anticarcinogenic and anti-inflammatory activities (Khan et al. 2015; Rumiyati et al. 2013). Phenolic compounds were characterized to have health benefits because of their high antioxidant capacity and protection against highly prevalent diseases (Van Hung 2016).

The fermentation of legumes enriched food products with high value proteins improved protein digestibility, changed an amino acid profile, reduced the concentration of antinutritional factors, increases antioxidant activity, thus improves nutritional characteristics (Yabaya et al. 2009; Curiel et al. 2015). Lactic acid bacteria (LAB) are important in various fermentation processes, implying that they have been proven to be safe for human consumption. Lactobacilli and pediococci prevent growth of spoilage and pathogenic microorganisms by acidification and production of antimicrobial compounds, contributing to improved food safety and quality (Cizeikiene et al. 2013).

Solid state fermentation (SSF) presents less energy requirements, lower potential of bacterial contamination, simple technology and equipment for fermentation compared to submerged fermentation (SmF) (Pandey et al. 2000).

Our previous studies showed, that fermentation improved nutritional properties of legume protein (Bartkiene et al. 2015). Diets to Wistar rats supplemented with fermented lupin products improved their gut environment (Bartkiene et al. 2013a). The use of fermented lupin flour additives reduce the acrylamide content in baked goods (Bartkiene et al. 2011, 2013b).

The object of this study was to evaluate the effect of submerged (SMF) and solid state fermentation (SSF) with Lactobacillus sakei KTU05-6 strain on the amino acids (AA) profile, total phenolics (TPC) content, and antioxidant activity of different Lupinus angustifolius seeds. Additionally, the impact of different lupin as fermentation media for LAB biomass and d/l-lactic acid production was analysed.

Materials and methods

Materials and microorganisms

Seeds of narrow-leaved lupin (Lupinus angustifolius L.) var. ‘Vilniai’ and six new hybrid lines (Nos. 1700, 1701, 1703, 1072, 1734, 1800) were obtained from the Voke Branch of Lithuanian Institute of Agriculture (Voke, Lithuania) in 2015.

The LAB strain Lactobacillus sakei KTU05-6, previously isolated from spontaneous rye sourdoughs (Digaitiene et al. 2012), were cultured at 30 °C temperature for 48 h in MRS medium (CM0359, Oxoid, Hampshire, United Kingdom) prior to be used.

Fermentation of lupin material

Lupin seeds were ground in a laboratory mill and obtained wholemeal was mixed with a LAB suspension, containing 8.9 log10 colony-forming units (cfu) per mL, followed by a fermentation in an anaerobic chamber (LabXMedia Group, Midland, Canada). The water content of samples was calculated with a reference to the moisture content of raw material and the required humidity of the end product [45% moisture content for solid state fermentation (SSF) and 65% for submerged fermentation (SMF)] Fermentation was carried out at 30 °C for 48 h. Each sample was analysed in triplicate for acidity, d- and l-lactic acid content and bacteria count. Unfermented lupin sample was analysed as a control.

Determination of acidity parameters

The pH values were measured and recorded with a pH electrode (Sartorius, Gottingen, Germany). The total titratable acidity (TTA) was determined by homogenizing a 10 g sample with 90 mL of distilled water and expressed as milliliters of 1 mol L−1 NaOH solution used to neutralise organic acids in 100 g of sample using the phenolphthalein as the indicator. The l(+)/d(−)-lactic acid concentrations in lupin samples were determined by an enzyme test kit (R-biopharm AG-Roche, Darmstadt, Germany).

Microbiological analysis

Ten grams of sample were homogenised with 90 mL of 9 g L−1 NaCl solution in distilled water. Samples after serial dilutions of 10−4–10−8 were sowed on sterile MRS agar (CM0361, Oxoid) on Petri plates. Samples were incubated under anaerobic conditions at 30 °C for 72 h. The number of LAB colonies was calculated and expressed as a log10 cfu g−1 of sample. All analysis were carried out in triplicate.

Preparation of lupin extracts

Lupin wholemeal (10 g) was transferred to dark-coloured flasks and mixed with 200 mL of methanol and stored at room temperature. After 24 h, infusions were filtered through Whatman No. 1 filter paper and residue was re-extracted with equal volume of solvent. Combined supernatants were evaporated to dryness under vacuum at 40 °C using Rotary evaporator. The obtained extracts were kept in sterile sample tubes and stored in a refrigerator at 4 ± 1 °C.

Determination of phenolic compounds

The total content of phenolic compounds (TPC) were determined by spectrophotometric method (Vaher et al. 2010). Methanolic solution of the sample extract in the concentration of 1 mg mL−1 was used in the analysis. The reaction mixture was prepared by mixing 0.5 mL of methanolic lupin extract, 2.5 mL of 10% Folin–Ciocalteu’s reagent dissolved in water and 2.5 mL 7.5% NaHCO3. Blank sample contained 0.5 mL methanol, 2.5 mL 10% Folin–Ciocalteu’s reagent dissolved in a water and 2.5 mL of 7.5% of NaHCO3. The samples were incubated at 45 °C for 40 min. The absorbance was measured at 765 nm using UV–Vis spectrophotometer (Genesys 10, Thermo Fisher Scientific Inc., Germany). The samples were prepared in triplicate for each analysis and the mean value of absorbance was obtained. The concentration of phenolics (mg mL−1) was calculated from the calibration curve, and the content of phenolics in extracts was expressed as gallic acid equivalent (mg of GA g−1 of extract).

Antioxidant activity determination

The ability of the lupin extract to scavenge DPPH free radicals was assessed by the standard method (Zhu et al. 2011). The stock solutions of extracts (1 mg/mL) were prepared in methanol. This solution (1 mL) was added to 3 mL of each extract in methanol at different concentrations (125, 250, 500 and 1000 μg mL−1). The mixtures were shaken vigorously and allowed to stand at room temperature for 30 min. Then the absorbance was measured at 517 nm using a UV–Vis spectrophotometer (Genesys 10, Thermo Fisher Scientific Inc., Germany) α-Tocopherol was used as the reference. The capability of scavenging the DPPH free radical was calculated by the following formula: DPPH scavenging effect (% inhibition) = (A0 − A1) × 100/A0 where, A0 is the absorbance of the control reaction, and A1 is the absorbance in presence of all of the extract samples and reference.

Determination of the amino acids content

Amino acids (AA) contents in lupin samples were analysed by gas chromatography (GC) with flame ionization detection after the ion-exchange solid phase extraction and chloroformate derivatization using EZ-Fast technology (Phenomenex, Torrance, CA, USA). Standard solutions of the amino acids alanine (ALA), glycine (GLY), valine (VAL), leucine (LEU), isoleucine (ILE), threonine (THR), serine (SER), proline (PRO), asparagine (ASP), methionine (MET), glutamine (GLU), phenylalanine (PHE), lysine (LYS), histidine (HIS), and tyrosine (TYR) were analysed, in addition to the internal standard (NVAL). Samples (1.00 g) were weighed in 15 mL polypropylene test tubes with screw caps and mixed with 7.5 mL of 0.1 M HCl, and subjected to ultrasonification in a water bath (t = 40 °C) for 15 min. The mixture was shaked and then centrifuged (3000 g, 15 min). An 2.5 mL aliquot of the mixture was transferred into another 15 mL polypropylene screw cap test tube and 7.5 mL of deionized water was added to 10 mL volume. Samples were then stored at −80 °C until analysis. The derivatized AA were analysed using a GC-FID instrument (6890 N, Agilent Technologies Inc., USA) equipped with an auto-sampler (7683 Series, Agilent Technologies Inc., USA). Aliquots of the derivatized amino acids (2 μL) were injected (1:15 split ratio) at 250 °C into a Zebron column (ZB-AAA, 10 m × 0.25 mm). Five different standard solutions with different concentrations (from 50 to 200 nmol μL−1) of AA standards were used for the calibration of gas chromatograph.

Statistical analysis

All analytical experiments were carried out in triplicate. In order to evaluate an influence of different factors (fermentation conditions and lupin variety) on acidity parameters, LAB count, AA profile, TPC content and antioxidant activity, data were subjected to analysis of variance (ANOVA) using statistical package SPSS for Windows (Ver.15.0, SPSS). The calculated mean values were compared using Tukey multiple range test with significance defined at p ≤ 0.05.

Results and disscussion

Effect of lupin seeds fermentation

Acidity parameters and viable LAB counts in lupin samples after SmF and SSF are presented in Table 1. The pH values in SmF and SSF lupin samples ranged from 3.67 to 3.95 and from 3.99 to 4.84. In all by SmF treated lupin samples pH was lower (8.5 to 9.3%) compared to SSF. The total titratable acidity (TTA) of fermented lupin ranged from 8.14 to 11.05 for SmF samples, and from 9.97 to 12.92 for SSF samples.

Table 1.

pH, total titratable acidity (TTA) values, d- and l-lactic acid contents (g 100 g−1 d.w.), and LAB counts (log10 cfu g−1) in L. angustifolius seeds after fermentation (48 h) with L. sakei

Samples ‘Vilniai’ No. 1072 No. 1734 No. 1700 No. 1701 No. 1800 No. 1702
pH
 SmF 3.90 ± 0.03b 3.67 ± 0.04a 3.64 ± 0.01a 3.70 ± 0.02a 3.72 ± 0.02a 3.67 ± 0.01a 3.76 ± 0.02a
 SSF 4.02 ± 0.03b 4.02 ± 0.02b 4.41 ± 0.03c 4.84 ± 0.03 4.20 ± 0.02b 4.37 ± 0.03bc 3.99 ± 0.02b
TTA
 SmF 8.75 ± 0.03b 8.89 ± 0.04b 8.14 ± 0.03a 11.05 ± 0.06d 9.82 ± 0.05c 11.04 ± 0.08d 9.52 ± 0.07c
 SSF 12.34 ± 0.05e 11.18 ± 0.05d 9.97 ± 0.04c 12.62 ± 0.11ef 10.80 ± 0.08 11.20 ± 0.08d 11.13 ± 0.08d
l(+)-lactate
 SmF 7.11 ± 0.10c 6.87 ± 0.27c 7.04 ± 0.22c 8.01 ± 0.15e 6.20 ± 0.20a 7.87 ± 0.06e 6.88 ± 0.22c
 SSF 7.52 ± 0.14d 7.61 ± 0.15d 7.88 ± 0.12e 9.01 ± 0.14g 6.41 ± 0.07b 8.27 ± 0.05f 7.67 ± 0.26d
d(−)-lactate
 SmF 1.14 ± 0.09a 3.12 ± 0.10 k 1.32 ± 0.19c 2.40 ± 0.14f 2.01 ± 0.22e 1.21 ± 0.18b 2.29 ± 0.12f
 SSF 2.87 ± 0.15g 1.41 ± 0.18d 1.23 ± 0.18b 1.09 ± 0.15a 2.97 ± 0.26h 2.89 ± 0.11g 2.88 ± 0.09g
LAB count
 SmF 9.12 ± 0.04a 9.21 ± 0.03a 9.38 ± 0.04a 9.78 ± 0.02c 9.46 ± 0.03b 9.00 ± 0.03a 9.90 ± 0.02c
 SSF 9.18 ± 0.03a 9.44 ± 0.02b 9.43 ± 0.04a 9.15 ± 0.03a 10.02 ± 0.04c 10.02 ± 0.02c 10.04 ± 0.02c
Open in a new tab

Data expressed as mean values (n = 3) ± SD

LAB lactic acid bacteria; SmF submerged fermentation; SSF solid state fermentation; SD standard deviation

a–hValues in the same row followed by different letters differ by Tukey test with significance defined at p ≤ 0.05

The viable cell number of tested LAB (in 48 h lupin fermentation) ranged from 9.0 to 9.9 log10 cfu g−1 in SmF samples and from 9.15 to 10.04 log10 cfu g−1 in SSF samples (Table 1). Because the growth of LAB in the fermentable substrate was limited by pH, the selection of substrates with strong buffer capacity allowed the production of products with high concentrations of organic acids and correspondingly high TTA values. In our study, there was not significant correlation obtained between the LAB cell count and pH, and between the LAB cells and TTA (r = 0.0134 and r = 0.0117, respectively) in the fermented lupin samples.

The TTA and pH are the most important parameters during fermentation processes (Yin et al. 2015). The LAB are a large group of closely related bacteria that have similar properties such as lactic acid production, which is an end product of the fermentation (Opere et al. 2012). Lactic acid produced by microbial fermentation usually is a mixture of l- and d-lactic acid. l-lactic acid was easily metabolized by humans, whereas d-lactic acid could be harmful when present in food in high concentrations (Reddy et al. 2008). According to Bartkiene et al. (2015), Lactobacillus sakei and Pediococcus pentosaceus strains produced mainly l-lactic acid in soya bean and lupin (d/l ratio 0.38–0.42 and 0.35–0.54, respectively), while spontaneous fermentation gave almost equal amounts of both lactic acid isomers (d/l ratio 0.82–0.98 and 0.92, respectively). In our study, the d/l ratio in fermented lupin varied from 0.15 to 0.45 (SmF) and from 0.12 to 0.46 (SSF), thus the L. sakei used for lupin fermentation could be indicated as a l-lactic acid producing bacteria.

Results of ANOVA test indicated a significant effect of lupin variety on TTA, l-lactate content, and LAB cell count. Also, it was found a significant influence of fermentation conditions (SmF and SSF) on substrate pH, TTA, and l-lactic acid concentration in lupin.

Total phenolic compounds and antioxidant activity of fermented lupin

Table 2 presents the total phenolic compounds (TPC) contents and antioxidant activity of fermented lupin. The TPC in unfermented lupin samples ranged from 318.21 (No. 1072) to 557.82 mg 100 g−1 d.w. (No. 1800). Fermented lupin have significantly (p < 0.05) higher TPC content than the unfermented seeds. Nutritional analysis highlighted a relevant increase in TPC content from 13.8–24.5% (No. 1702, No. 1800, ‘Vilniai’ and No. 1700) to 39.0–65.2% (No. 1734, No. 1701 and No. 1072) in by SSF treated L. angustifolius seeds, compared to unfermented samples. ANOVA test indicated a significant effect of lupin variety and fermentation type (SmF and SSF) on TPC content in lupin seeds.

Table 2.

Total phenolic compounds (TPC) (mg 100 g−1 d.w.) and antioxidant activity (%) of L. angustifolius seeds before and after fermentation with L. sakei

Samples ‘Vilniai’ No. 1072 No. 1734 No. 1701 No. 1700 No. 1800 No. 1702
TPC
 Control 500.38 ± 4.89d 318.21 ± 3.54a 466.31 ± 4.58d 408.87 ± 3.79c 419.29 ± 3.86c 557.82 ± 5.22f 552.47 ± 5.16f
 SmF 510.61 ± 3.49d 382.69 ± 4.02b 513.05 ± 4.01d 469.69 ± 4.82d 401.82 ± 3.60bc 553.88 ± 5.20f 524.31 ± 5.03e
 SSF 586.54 ± 5.32g 525.32 ± 5.16e 648.20 ± 6.15j 596.95 ± 5.34g 522.06 ± 4.93e 650.54 ± 5.00j 628.58 ± 4.77h
Antioxidant activity (DPPH)
 Control 59.28 ± 0.98a 61.04 ±  ± 0.97a 68.51 ± 1.02c 64.54 ± 0.97b 70.27 ± 1.09d 74.24 ± 1.16d 72.21 ± 1.12c
 SmF 65.65 ± 1.01b 62.98 ± 0.98ab 66.85 ± 1.04c 74.15 ± 1.16d 84.12 ± 1.36e 83.29 ± 1.34e 75.62 ± 1.14d
 SSF 88.18 ± 1.41f 65.37 ± 0.52b 74.24 ± 0.97d 71.19 ± 1.08c 71.93 ± 1.10c 62.05 ± 0.88a 83.65 ± 1.31e
Open in a new tab

Data expressed as mean values (n = 3) ± SD

Control—unfermented lupin. SmF submerged fermentation; SSF solid state fermentation; SD standard deviation

a–jValues in the same row followed by different letters differ by Tukey test with significance defined at p ≤ 0.05

The results of analysis of antioxidant activity showed that unfermented lupin have antioxidant activity varying between 59.28 and 74.24% in L. angustifolius seeds. The increase in antioxidant activity after 48 h fermentation was noticed in all lupins ranging from 62.98 (No. 1072) to 84.12% (No. 1700) in SmF samples, and from 62.05 (No. 1800) to 88.18% (‘Vilniai’) in SSF samples. There was found a significant effect of lupin variety on the antioxidant activity, however the significant impact of fermentation type (SmF and SSF) on the antioxidant activity of lupin seeds was not observed. Furthermore, there was not a significant relation between the TPC and antioxidant activity of fermented lupin.

Fermented lupin showed higher antioxidant activities compared to unfermented lupin at the concentrations tested. In the literature has been reported that fermentation enhances micro nutrient bioavailability and aids in degrading antinutritional factors (Oboh and Rocha 2007). Fermentation also improved the phenolic content and antioxidant properties of legume seeds (Ademiluyi and Oboh 2011). Since lupin seeds have been found to be of high protein content, the breakdown of protein to free amino acids and peptides by microbial protease activity could also increase the antioxidant activity. Watanabe et al. (2007) after modification of the fermentation conditions observed that isoflavoneaglycones, free amino acids and peptides were responsible for the antioxidant activity of tempeh. Basically, the improvement in antioxidant activity occurs when microorganisms start breaking down the linkage of phenolic and flavonoid compounds, which free radicals actively play the role of antioxidants (Moktan et al. 2008; Ademiluyi and Oboh 2011).

Amino acids profile in fermented and nonfermented lupin

Tables 3 and 4 present the EAA and NEAA content of unfermented and fermented with L. sakei lupin, respectively. Among all EAA, the LEU was found to be of the highest content while MET (0.15–1.38% from total AA) was the one of lowest content in all lupins (Table 3). The results showed that L. angustifolius seeds contained the highest amounts of LEU (7.05–9.34% from TTA), LYS (3.63–7.66% from TAA), PHE (4.75–6.41% from TAA) depending on lupin variety (Table 3). Our results were similar to Iqbal et al. (2006) which reported that legumes were deficient in amino acids such as methionine, cystine and cysteine.

Table 3.

Essential amino acids (EAA) content in percentage from the total amino acids in L. angustifolius seeds before and after fermentation with L. sakei

Samples VAL ILE LEU THR MET PHE LYS HIS
Unfermented (controls)
 ‘Vilniai’ 4.32 ± 0.05a 4.83 ± 0.04a 7.05 ± 0.04a 3.45 ± 0.04a 0.15 ± 0.01a 4.75 ± 0.03a 7.16 ± 0.09d 4.34 ± 0.05a
 No. 1072 4.56 ± 0.05b 5.10 ± 0.05b 7.39 ± 0.04b 3.75 ± 0.03b 0.15 ± 0.01a 5.10 ± 0.05b 7.66 ± 0.09e 4.40 ± 0.05a
 No. 1734 4.69 ± 0.06b 5.12 ± 0.03b 8.43 ± 0.06c 4.26 ± 0.04c 0.17 ± 0.02c 5.58 ± 0.05c 5.83 ± 0.06b 4.30 ± 0.04a
 No. 1701 4.74 ± 0.04c 5.05 ± 0.05b 7.36 ± 0.04a 3.87 ± 0.03b 0.15 ± 0.01a 5.06 ± 0.05b 6.40 ± 0.06c 4.47 ± 0.05a
 No. 1700 6.07 ± 0.08d 6.04 ± 0.07c 9.34 ± 0.07a 4.36 ± 0.05c 1.38 ± 0.02d 6.41 ± 0.06d 3.63 ± 0.04a 9.27 ± 0.05d
 No. 1800 4.50 ± 0.04a 4.89 ± 0.04a 7.32 ± 0.05a 3.76 ± 0.03b 0.17 ± 0.01c 4.85 ± 0.05a 6.51 ± 0.07c 4.37 ± 0.04a
 No. 1702 4.54 ± 0.04b 5.09 ± 0.05b 7.47 ± 0.05b 3.69 ± 0.03b 0.15 ± 0.01a 5.17 ± 0.06b 6.47 ± 0.07c 4.78 ± 0.05b
Fermented (SmF)
 ‘Vilniai’ 6.50 ± 0.04c 4.39 ± 0.04a 8.11 ± 0.05c 6.33 ± 0.02d 2.08 ± 0.0e 4.62 ± 0.05a 7.95 ± 0.08g 4.18 ± 0.04d
 No. 1072 6.48 ± 0.04c 4.53 ± 0.04a 7.50 ± 0.05b 6.14 ± 0.02d 1.58 ± 0.03c 4.77 ± 0.04b 6.39 ± 0.07f 3.97 ± 0.03d
 No. 1734 6.75 ± 0.05d 4.64 ± 0.04b 7.52 ± 0.05b 6.39 ± 0.04d 1.76 ± 0.01d 4.53 ± 0.04a 5.24 ± 0.05d 3.69 ± 0.03c
 No. 1701 8.76 ± 0.06f 5.69 ± 0.06e 8.64 ± 0.06d 4.15 ± 0.02b 1.43 ± 0.02b 5.63 ± 0.06d 4.90 ± 0.05c 4.54 ± 0.04e
 No. 1700 5.22 ± 0.05a 9.34 ± 0.08g 6.04 ± 0.06a 4.36 ± 0.04c 0.94 ± 0.01a 6.41 ± 0.06e 3.63 ± 0.03a 4.84 ± 0.04f
 No. 1800 7.68 ± 0.05e 6.43 ± 0.07f 9.67 ± 0.08e 3.77 ± 0.04a 1.97 ± 0.02e 6.59 ± 0.07e 5.73 ± 0.06e 5.73 ± 0.06g
 No. 1702 8.42 ± 0.05f 5.33 ± 0.06d 8.79 ± 0.06d 4.30 ± 0.04b 1.62 ± 0.04c 5.32 ± 0.05c 4.20 ± 0.04b 2.87 ± 0.02a
Fermented (SSF)
 ‘Vilniai’ 5.32 ± 0.03d 4.65 ± 0.04a 9.42 ± 0.08c 6.54 ± 0.04f 0.81 ± 0.05b 5.92 ± 0.06b 6.10 ± 0.04e 5.75 ± 0.07d
 No. 1072 4.83 ± 0.05b 5.39 ± 0.05b 9.32 ± 0.08b 5.83 ± 0.05e 0.90 ± 0.04c 6.55 ± 0.07d 4.29 ± 0.04b 6.36 ± 0.07e
 No. 1734 4.79 ± 0.05b 5.28 ± 0.05b 8.30 ± 0.07a 4.23 ± 0.04b 0.84 ± 0.04b 5.31 ± 0.05a 3.99 ± 0.03a 4.90 ± 0.04c
 No. 1701 5.08 ± 0.05c 6.15 ± 0.07d 15.41 ± 0.06e 3.98 ± 0.03a 0.89 ± 0.03c 6.92 ± 0.07e 4.31 ± 0.04b 4.75 ± 0.04c
 No. 1700 6.83 ± 0.07e 5.74 ± 0.06c 11.78 ± 0.08d 5.50 ± 0.04d 1.54 ± 0.04e 5.81 ± 0.06b 5.31 ± 0.04d 3.36 ± 0.02a
 No. 1800 5.01 ± 0.05c 5.66 ± 0.06c 8.92 ± 0.07b 5.51 ± 0.04d 0.67 ± 0.04a 6.07 ± 0.06c 4.30 ± 0.03b 3.44 ± 0.03a
 No. 1702 5.39 ± 0.05d 5.03 ± 0.04a 8.19 ± 0.06a 5.28 ± 0.05c 1.07 ± 0.05d 5.83 ± 0.06b 4.76 ± 0.04c 3.34 ± 0.03a
Open in a new tab

Data expressed as mean values (n = 3) ± SD

SmF submerged fermentation; SSF solid state fermentation; SD standard deviation

a–gValues in the same column followed by different letters differ by Tukey test with significance defined at p ≤ 0.05

Table 4.

Non-essential amino acids (NEAA) content in percentage from the total amino acids in L. angustifolius seeds before and after fermentation with L. sakei

Samples ALA GLY SER PRO ASP GLU TYR
Unfermented (controls)
 ‘Vilniai’ 3.14 ± 0.04a 3.83 ± 0.04a 5.02 ± 0.04a 4.35 ± 0.04a 10.43 ± 0.10a 26.58 ± 0.16b 10.61 ± 0.10e
 No. 1072 3.37 ± 0.03c 4.06 ± 0.04b 5.53 ± 0.06c 4.67 ± 0.05b 11.34 ± 0.12b 27.34 ± 0.15c 5.57 ± 0.04b
 No. 1734 3.99 ± 0.04e 4.59 ± 0.05d 6.11 ± 0.07d 4.70 ± 0.06b 10.39 ± 0.11a 26.20 ± 0.17b 5.64 ± 0.06b
 No. 1701 3.26 ± 0.03b 4.19 ± 0.04b 5.72 ± 0.05c 4.72 ± 0.04b 11.77 ± 0.12c 24.86 ± 0.16a 8.40 ± 0.07d
 No. 1700 3.64 ± 0.02d 4.38 ± 0.05c 5.90 ± 0.06d 5.08 ± 0.05c 12.16 ± 0.13d 26.25 ± 0.18b 3.58 ± 0.02a
 No. 1800 3.27 ± 0.03b 3.99 ± 0.04b 5.63 ± 0.06c 4.53 ± 0.05b 11.31 ± 0.11b 27.80 ± 0.17c 7.00 ± 0.06c
 No. 1702 3.22 ± 0.04b 4.05 ± 0.05b 5.38 ± 0.04b 4.69 ± 0.04b 11.04 ± 0.12a 27.54 ± 0.19c 6.74 ± 0.05c
Fermented (SmF)
 ‘Vilniai’ 3.58 ± 0.03a 3.98 ± 0.03a 7.29 ± 0.07f 4.56 ± 0.04a 11.37 ± 0.12b 30.43 ± 0.18e 2.32 ± 0.02a
 No. 1072 3.37 ± 0.03a 4.06 ± 0.04a 5.53 ± 0.05a 4.67 ± 0.05a 11.34 ± 0.10b 27.34 ± 0.16d 5.57 ± 0.05e
 No. 1734 3.99 ± 0.04b 4.59 ± 0.05c 6.11 ± 0.06b 4.70 ± 0.04a 10.39 ± 0.11a 26.20 ± 0.15d 5.64 ± 0.05e
 No. 1701 4.79 ± 0.05d 5.36 ± 0.06e 6.96 ± 0.07e 5.56 ± 0.06d 12.61 ± 0.13c 21.40 ± 0.13b 3.97 ± 0.03b
 No. 1700 4.18 ± 0.04c 5.27 ± 0.05e 6.65 ± 0.06d 6.43 ± 0.07f 12.95 ± 0.14c 19.85 ± 0.17a 3.88 ± 0.04b
 No. 1800 3.80 ± 0.04b 4.63 ± 0.05c 6.32 ± 0.06c 5.95 ± 0.07e 11.45 ± 0.11b 19.36 ± 0.16a 4.71 ± 0.04c
 No. 1702 4.63 ± 0.05d 4.87 ± 0.04d 6.55 ± 0.06d 5.37 ± 0.06c 12.86 ± 0.12c 23.76 ± 0.18c 5.23 ± 0.05d
Fermented (SSF)
 ‘Vilniai’ 3.91 ± 0.02b 5.39 ± 0.05c 7.23 ± 0.06b 5.85 ± 0.05a 12.12 ± 0.11a 23.38 ± 0.14d 1.97 ± 0.01a
 No. 1072 3.88 ± 0.03b 5.75 ± 0.06d 7.12 ± 0.05b 6.76 ± 0.06c 13.33 ± 0.13b 18.57 ± 0.17a 2.37 ± 0.02c
 No. 1734 3.82 ± 0.04b 5.09 ± 0.05a 7.67 ± 0.07c 5.78 ± 0.05a 13.04 ± 0.12b 25.43 ± 0.18e 2.11 ± 0.01b
 No. 1701 3.40 ± 0.02a 5.07 ± 0.04a 6.18 ± 0.06a 6.67 ± 0.07c 12.89 ± 0.11b 20.53 ± 0.19b 3.87 ± 0.03d
 No. 1700 4.25 ± 0.04c 4.84 ± 0.03a 7.27 ± 0.07b 5.78 ± 0.05a 13.60 ± 0.13c 19.28 ± 0.17a 6.50 ± 0.06f
 No. 1800 3.79 ± 0.03b 4.91 ± 0.04a 6.40 ± 0.05a 6.15 ± 0.05b 13.53 ± 0.13c 21.91 ± 0.15c 4.93 ± 0.04e
 No. 1702 4.27 ± 0.04c 5.16 ± 0.05b 7.68 ± 0.06c 6.24 ± 0.06b 12.43 ± 0.11a 19.23 ± 0.14a 7.50 ± 0.05g
Open in a new tab

Data expressed as mean values (n = 3) ± SD

SmF submerged fermentation; SSF solid state fermentation; SD standard deviation

a–gValues in the same column followed by different letters differ by Tukey test with significance defined at p ≤ 0.05

The use of the SmF as well as SSF significantly increased the EAA contents in L. angustifolius samples due to increased VAL [6.48–8.76% (SmF) and 4.79–6.83% (SSF) from TAA], LEU [7.50–9.67% (SmE) and 8.19–15.41% (SSF) from TAA], MET (0.94–2.08% (SmE) and 0.67–1.54% (SSF) from TAA), THR [4.36–6.39% (SmE) and 3.98–6.54% (SSF) from TAA] contents with the exeption of hybrid No. 1700.

The amounts of other EAA were found to vary among the lupin varieties. Statistical analysis showed a significant effect of lupin variety on different EAA contents.

Furthermore, data analysis showed a significant effect of fermentation conditions on EAA content in lupin. Either, the significant interaction of these factors (seed variety and fermentation conditions) was obtained on total EAA content in lupin. The hydrolytic breakdown of the nutrient components during fermentation may have caused the increase in AA content (Ferial and Esmat 2011).

The amount of remaining non-essential amino acids (NEAA) varied depending on lupin variety. The amino acids such as ALA, followed by GLY and PRO was found to be in lowest contents (Table 4). The results indicate that in most cases (aprox. 50% of samples) the SSF reduces the NEAA contents (Table 4), while the SmF conditions has lower impact on changes in the NEAA profile. The reduction in NEAA content on average by 6.6% was observed in L. angustifolius var. ‚Vilniai‘and hybrids No. 1072, 1734 and 1701 after SSF, and by 6.1% in hybrids Nos. 1701, 1700 and 1800 after the SmF due to lower contents of TYR (from 3.58–10.61 to 1.97–7.5%) and GLU (24.86–27.80 to 18.57–30.43%) compared to unfermented lupin.

The SmF as well as SSF with L. sakei strain increased the ALA content in all lupin samples (from 2.92–3.99 to 3.37–4.79%) with the exception of lupin No. 1734. Also, in fermented lupin samples higher contents of GLY (from 3.79–4.59 to 3.98–5.75%), SER (5.02–6.11 to 5.53–7.68%), PRO (4.35–5.08 to 4.56–6.76%), and ASP (10.39–12.16 to 10.39–13.60%) were determined compared to controls.

Results of ANOVA indicated a significant effect of lupin variety on NEAA content (p ≤ 0.05), with the exception of GLY, SER and TYR. Fermentation type had a significant influence on all NEAA contents (p < 0.0001), and statistically significant interaction of this factors on the NEAA in lupin was observed. The NEAA should be taken into consideration in revising the ideal protein concept and formulating balanced diets to improve protein accretion, food nutritional value, and human health (Wu et al. 2013).

Conclusion

The present study represent amino acids profile and antioxidant activity of seeds of L. angustifolius a wild plant of Baltic Sea region. The results demonstrated that fermentation caused a marked increase in total phenolic contents and antioxidant activity of lupin which enhanced DPPH radical-scavenging ability. Fermentation for 48 h was applicable as exemplified by the DPPH radical-scavenging ability of the lupin seeds. The interaction of the lupin variety and fermentation conditions had a significant effect on the EAA and NEAA contents. Considering total phenolics, DPPH radical-scavenging ability and improved EAA profile, fermented lupin showed greater nutritional quality compared to unfermented.

Contributor Information

Vytaute Starkute, Email: vytaute.starkute@lsmuni.lt.

Elena Bartkiene, Phone: +370 37 574565, Email: Elena.Bartkiene@lsmuni.lt.

Vadims Bartkevics, Email: vadims.bartkevics@bior.gov.lv.

Janis Rusko, Email: janis.rusko@gmail.com.

Daiva Zadeike, Email: daiva.zadeike@ktu.lt.

Grazina Juodeikiene, Email: grazina.juodeikiene@ktu.lt.

References

  1. Abdelrahman AR. Influence of chemical properties of wheat–lupine flour blends on cake quality. Am J Food Sci Technol. 2014;2:67–75. doi: 10.12691/ajfst-2-2-4. [DOI] [Google Scholar]
  2. Ademiluyi AO, Oboh G. Antioxidant properties of condiment produced from fermented bambara groundnut (Vigna subterranean L. Verdc) J Food Biochem. 2011;35:1145–1160. doi: 10.1111/j.1745-4514.2010.00441.x. [DOI] [Google Scholar]
  3. Bähr M, Fechner A, Kiehntopf M, Jahreis G. Consuming a mixed diet enriched with lupin protein beneficially affects plasma lipids in hypercholesterolemic subjects: a randomized controlled trial. Clin Nutr. 2015;34:7–14. doi: 10.1016/j.clnu.2014.03.008. [DOI] [PubMed] [Google Scholar]
  4. Bartkiene E, Juodeikiene G, Vidmantiene D, Viskelis P, Urbonaviciene D. Nutritional and quality aspects of wheat sourdough bread using L. luteus and L. angustifolius flours fermented by Pedioccocus acidilactici. Int J Food Sci Technol. 2011;46:1724–1733. doi: 10.1111/j.1365-2621.2011.02668.x. [DOI] [Google Scholar]
  5. Bartkiene E, Juodeikiene G, Vidmantiene D, Zdunczyk Z, Zdunczyk P, Juśkiewicz J, Cizeikiene D, Matusevicius P. Influence of diets to Wistar rats supplemented with soya, flaxseed and lupine products treated by lactofermentation to improve their gut health. Int J Food Sci Nutr. 2013;64:730–739. doi: 10.3109/09637486.2013.775230. [DOI] [PubMed] [Google Scholar]
  6. Bartkiene E, Juodeikiene G, Vidmantiene D. Nutritional quality of fermented defatted soya and flaxseed flours and their effect on texture and sensory characteristics of wheat sourdough bread. Int J Food Sci Nutr. 2013;63:722–729. doi: 10.3109/09637486.2011.649248. [DOI] [PubMed] [Google Scholar]
  7. Bartkiene E, Krungleviciute V, Juodeikiene G, Vidmantiene D, Maknickiene Z. Solid state fermentation with lactic acid bacteria to improve the nutritional quality of lupin and soya bean. J Sci Food Agric. 2015;95:1336–1342. doi: 10.1002/jsfa.6827. [DOI] [PubMed] [Google Scholar]
  8. Carbonaro M, Maselli P, Nucara A. Structural aspects of legume proteins and nutraceutical properties. Food Res Int. 2015;76:19–30. doi: 10.1016/j.foodres.2014.11.007. [DOI] [Google Scholar]
  9. Cizeikiene D, Juodeikiene G, Paskevicius A, Bartkiene E. Antimicrobial activity of lactic acid bacteria against pathogenic and spoilage microorganism isolated from food and their control in wheat bread. Food Control. 2013;31:539–545. doi: 10.1016/j.foodcont.2012.12.004. [DOI] [Google Scholar]
  10. Curiel JA, Coda R, Centomani I, Summo C, Gobbetti M, Rizzello CG. Exploitation of the nutritional and functional characteristics of traditional Italian legumes: the potential of sourdough fermentation. Int J Food Microbiol. 2015;196:51–61. doi: 10.1016/j.ijfoodmicro.2014.11.032. [DOI] [PubMed] [Google Scholar]
  11. Digaitiene A, Hansen AS, Juodeikiene G, Eidukonyte D, Josephsen J. Lactic acid bacteria isolated from rye sourdouhs produce bacteriocin-like inhibitory substances active against Bacillus subtilis and fungi. J Appl Microbiol. 2012;112:732–742. doi: 10.1111/j.1365-2672.2012.05249.x. [DOI] [PubMed] [Google Scholar]
  12. Ferial MAS, Esmat AAA. Physico-chemical properties of tempeh produced from chickpea seeds. J Am Sci. 2011;7(7):107–118. [Google Scholar]
  13. Galili G. The aspartate-family pathway of plants: linking production of essential amino acids with energy and stress regulation. Plant Signal Behav. 2011;6:192–195. doi: 10.4161/psb.6.2.14425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Galili G, Amir R. Fortifying plants with the essential amino acids lysine and methionine to improve nutritional quality. Plant Biotechnol J. 2013;11:211–222. doi: 10.1111/pbi.12025. [DOI] [PubMed] [Google Scholar]
  15. Galili G, Amir R, Fernie AR. The regulation of essential amino acid synthesis and accumulation in plants. Annu Rev Plant Biol. 2016;67:153–178. doi: 10.1146/annurev-arplant-043015-112213. [DOI] [PubMed] [Google Scholar]
  16. Iqbal A, Khalil IA, Ateeq N, Khan SM. Nutritional quality of important food legumes. Food Chem. 2006;97(2):331–335. doi: 10.1016/j.foodchem.2005.05.011. [DOI] [Google Scholar]
  17. Jayasena V, Nasar-Abbas SM. Development and quality evaluation of high-protein and high-dietary fiber pasta using lupin flour. J Texture Stud. 2012;43:153–163. doi: 10.1111/j.1745-4603.2011.00326.x. [DOI] [Google Scholar]
  18. Khan MK, Karnpanit W, Nasar-Abbas SM, Hum ZE, Jayasena V. Phytochemical composition and bioactivities of lupin: a review. Int J Food Sci Technol. 2015;50:2004–2012. doi: 10.1111/ijfs.12796. [DOI] [Google Scholar]
  19. Moktan B, Saha J, Sarkar PK. Antioxidant activities of soybean as affected by Bacillus fermentation to kinema. Food Res Int. 2008;41:586–593. doi: 10.1016/j.foodres.2008.04.003. [DOI] [Google Scholar]
  20. Oboh G, Rocha JBT. Antioxidant in foods: a new challenge for food processors. In: Panglossi HV, editor. New development in antioxidants research. New York: Nova Science Publishers Inc; 2007. pp. 35–64. [Google Scholar]
  21. Opere BOO, Aboaba EO, Ugoji A, Iwalokun BA. Estimation of nutritive value, organoleptic properties and consumer acceptability of fermented cereal gruel (OGI) Adv J Food Sci Technol. 2012;4:1–8. [Google Scholar]
  22. Orlovskaya TV, Sultanova EM, Oshchepkova YuI, Arzanova IA, Kuznetsova NN, Veshkurova ON, Salikhov ShI. Bioactive peptides from several plant species. Chem Nat Compd. 2010;46(2):331–332. doi: 10.1007/s10600-010-9608-3. [DOI] [Google Scholar]
  23. Pandey A, Soccol CR, Mitchel D. New developments in solid state fermentation: I-bioprocesses and products. Process Biochem. 2000;35:1153–1169. doi: 10.1016/S0032-9592(00)00152-7. [DOI] [Google Scholar]
  24. Reddy G, Altaf MD, Naveena BJ, Venkateshwar M, Kumar EV. Amylolytic bacterial lactic acid fermentation—a review. Biotechnol Adv. 2008;26:22–34. doi: 10.1016/j.biotechadv.2007.07.004. [DOI] [PubMed] [Google Scholar]
  25. Rumiyati R, Jayasena V, James P. Total phenolic and phytosterol compounds and the radical scavening activity of germinated Australian sweet lupin flour. Plant Food Hum Nutr. 2013;68:352–357. doi: 10.1007/s11130-013-0377-6. [DOI] [PubMed] [Google Scholar]
  26. Rumiyati R, James AP, Jayasena V. Effects of lupin incorporation on the physical properties and stability of bioactive constituents in muffins. Int J Food Sci Technol. 2015;50:103–110. doi: 10.1111/ijfs.12688. [DOI] [Google Scholar]
  27. Thambiraj SR, Phillips M, Koyyalamudi SR, Reddy N. Antioxidant activities and characterisation of polysaccharides isolated from the seeds of Lupinus angustifolius. Ind Crops Prod. 2015;74:950–956. doi: 10.1016/j.indcrop.2015.06.028. [DOI] [Google Scholar]
  28. Vaher M, Matso K, Levandi T, Helmja K, Kaljurand M. Phenolic compounds and the antioxidant activity of the bran, flour and whole grain of different wheat varieties. Proc Chem. 2010;2:76–82. doi: 10.1016/j.proche.2009.12.013. [DOI] [Google Scholar]
  29. Van Hung P. Phenolic compounds of cereals and their antioxidant capacity. Crit Rev Food Sci Nutr. 2016;56:25–35. doi: 10.1080/10408398.2012.708909. [DOI] [PubMed] [Google Scholar]
  30. Villarino CBJ, Jayasena V, Coorey R, Chakrabarti-Bell S, Johnson SK. The effects of Australian sweet lupin (ASL) variety on physical properties of flours and breads. LWT Food Sci Technol. 2015;60:435–443. doi: 10.1016/j.lwt.2014.08.028. [DOI] [Google Scholar]
  31. Watanabe N, Fujimoto K, Aoki H. Antioxidant activities of the water-soluble fraction in tempeh-like fermented soybean (GABA-tempeh) Int J Food Sci Nutr. 2007;58:577–587. doi: 10.1080/09637480701343846. [DOI] [PubMed] [Google Scholar]
  32. Wu G, Wu Z, Dai Z, Yang Y, Wang W, Liu C, Wang B, Wang J, Yin Y. Dietary requirements of nutritionally non-essential amino acids by animals and humans. Amino Acids. 2013;44:1107–1113. doi: 10.1007/s00726-012-1444-2. [DOI] [PubMed] [Google Scholar]
  33. Yabaya A, Akinyanju JA, Jatou ED. Yeast enrichment of soybean cake. World J Dairy Food Sci. 2009;4:141–144. [Google Scholar]
  34. Yin Y, Jinshui W, Sen Y, Jingli F, Feng J, Changfu Z. Protein degradation in wheat sourdough fermentation with Lactobacillus plantarum M616. Interdiscip Sci. 2015;7:205–210. doi: 10.1007/s12539-015-0262-0. [DOI] [PubMed] [Google Scholar]
  35. Zhu KX, Lian CX, Guo XN, Peng W, Zhou HM. Antioxidant activities and total phenolic contents of various extracts from defatted wheat germ. Food Chem. 2011;126:1122–1126. doi: 10.1016/j.foodchem.2010.11.144. [DOI] [Google Scholar]

Articles from Journal of Food Science and Technology are provided here courtesy of Springer

RESOURCES