胁迫条件下褪黑素诱导雨生红球藻虾青素合成

雨生红球藻(Haematococcus pluvialis)在胁迫条件下可大量积累虾青素,已成为天然虾青素的主要来源。通过解析外源褪黑素(Melatonin,MLT)调控雨生红球藻在缺氮联合高光照胁迫条件下的防御效应,以期建立虾青素高效合成的技术体系。结果表明,胁迫条件下外源MLT的诱导显著促进了虾青素的积累,最高质量分数达到32.37 mg/g,较对照组增加了2.25倍。此外,外源MLT提高了胞内NO和丝裂原活化蛋白激酶(MAPK)的含量,同时上调了虾青素合成关键酶基因dxs和chy的表达水平。研究表明,外源MLT诱导高光缺氮胁迫下雨生红球藻中虾青素的高效合成可能与MLT调控藻细胞内的NO、MAPK含量和虾青素合成关键酶基因dxs和chy的表达水平相关。     

Supercritical carbon dioxide extraction of astaxanthin and other carotenoids from the microalga Haematococcus pluvialis

Supercritical carbon dioxide extraction of astaxanthin and other carotenoids from Haematococcus pluvialis was carried out, for several experimental conditions, using a semi-continuous apparatus. The microalga was previously freeze-dried and ground with a ball mill. The effects of pressure (200 and 300 bar), temperature (40 and 60 °C), degree of crushing, as well as the use of ethanol as a co-solvent (10%) on the extraction efficiency were assessed. Organic solvent extractions, using acetone, were also carried out in a vortex, on ground cells mixed with very small glass beads. Supercritical extraction from the completely crushed alga was compared with acetone and the highest recovery of carotenoids (92%) was obtained at the pressure of 300 bar and the temperature of 60 °C, using ethanol as a co-solvent.The extraction recovery increased with the pressure at 60 °C. On the other hand, the increase in temperature, at 300 bar, led to a slight improvement. The main carotenoid of Haematococcus pluvialis is the esterified astaxanthin (about 75%). Other carotenoids present are lutein, astaxanthin (free), β-carotene and canthaxanthin. All of them were recovered through supercritical fluid extraction with values higher than 90%, with the exception of canthaxanthin (about 85%), at a pressure of 300 bar and a temperature of 60 °C.     

Effects of nutrient supply methods and illumination with blue light emitting diodes (LEDs) on astaxanthin production by Haematococcus pluvialis

In order to increase the cell concentration and the accumulation of astaxanthin, the effects of nutrient concentration, pH, illumination and methods of supplying nutrients were studied for the cultivation of Haematococcus pluvialis. The replacement of media to avoid the deficiency of nutrients increased the cell concentration above 1 mg-dry cell cm(-3) without induction of astaxanthin accumulation. Illumination with blue light emitting diode lamps and nutrient starvation induced accumulation of astaxanthin, and the interactive effects of these two increased the astaxanthin concentration to 76 mug cm(-3).     

Effects of parameters affecting biomass yield and thermal behaviour of Chlorella vulgaris

Conventional fossil fuels are facing a global challenge which lead scientists to explore alternative fuel production from biological sources. The algae-based fuels are gaining rapid attention as it has potential to replace petroleum-based fuels. An indigenous high lipid producing microalgae was isolated from a freshwater pond in the KwaZulu-Natal province of South Africa. The isolate was later identified as Chlorella vulgaris, based on partial 28S large subunit ribosomal RNA gene sequence. The growth kinetics, pyrolytic characteristics and photosynthetic efficiency of Chlorella was evaluated in vitro. The optimized conditions for higher biomass yield of the selected strain were at 4% CO(2), 0.5 g l(-1) NO(3) and 0.04 g l(-1) PO(4), respectively. The pulse amplitude modulation results indicated that C. vulgaris could withstand a light intensity ranging from 150 to 350 μmol photons m(-2)s(-1). Further increase in light intensity resulted in a decline of the electron transport rate. Carbon fixation rate, lipid content and calorific value of C. vulgaris was 6.17 mg l(-1)h(-1), 21% and 17.44 kJ g(-1), respectively. The pyrolitic studies under inert atmosphere at different heating rates of 15, 30, 40 and 50°C min(-1) from ambient temperature to 800°C showed that the overall final weight loss recorded for the four different heating rates was in the range of 78.9-81%. These studies could be useful to appraise the biofuel potential of the isolated C. vulgaris strain, which can later be taken for pilot scale production.     

Amino acid,structure and antioxidant properties of Haematococcus pluvialis protein hydrolysates produced by different proteases

The impact of different protease hydrolysis on the amino acid, structure and antioxidant properties of H. pluvialis protein (HP) was investigated. Results showed that the hydrolysate obtained by Alcalase exhibited the highest degree of hydrolysis (20.59%) and peptide yield (92.64%). The essential amino acid, hydrophobic, sulphur and aromatic amino acid contents of enzyme hydrolysates were significantly higher than HP (P < 0.05). FTIR spectra showed that the β-sheet proportion of HP hydrolysates were higher compared with HP, the proportion of random coil structure was lower. The α-helix content of the hydrolysate obtained by Alcalase was the highest, while the turn proportion was the lowest. The Trypsin derived hydrolysate presented the best DPPH and ABTS scavenging ability, and ferric reducing antioxidant power than other HPHs. These results suggested that HP hydrolysates have a great potential as natural functional ingredients in food manufacture.     

Pressurized liquids as an alternative process to antioxidant carotenoids' extraction from Haematococcus pluvialis microalgae

In this work, extraction of antioxidant carotenoids from Haematococcus pluvialis microalga, has been studied combining pressurized liquid extraction (PLE), using hexane and ethanol as extracting solvents, and analytical techniques such as thin layer chromatography (TLC) and HPLC with DAD. The effect of the extraction temperature (50, 100, 150 and 200 °C) and the polarity of the solvent have been studied in terms of in vitro antioxidant activity and chemical composition considering two different morphological cells (green vegetative cells and red cysts). Results demonstrate that the extraction temperature had a positive influence in the extraction yield while its effect in the antioxidant activity was negative, lowering the activity of the extracts with an increase of the extraction temperature. The best yields were obtained with ethanol at the higher extraction temperature while the best antioxidant activity was also achieved using ethanol but at lower temperatures. Chemical composition was determined by TLC and HPLC with DAD. Several compounds were identified in the samples and concentration of astaxanthin was obtained. Results pointed out that the extracts contained different carotenoids in both, the green and the red phase, and that depending on its contribution a stronger antioxidant activity would be expected.     

Enlarged and astaxanthin-accumulating cyst cells of the green alga Haematococcus pluvialis

The cyst cells of Haematococcus pluvialis were separated into fractions of relatively uniform size by sucrose density gradient centrifugation. The fraction at the bottom of the centrifuge tube with the largest specific gravity from density gradients of mature cysts mainly consisted of enlarged, red cyst cells and had the highest astaxanthin content. To examine the relationship between cell size and astaxanthin content of cysts, formation of the fluorescent dichlorofluorescein (DCF) from 2′,7′-dichlorohydrofluorescein diacetate of cyst cells in each fraction from density-gradient centrifugation under oxidative stress caused by methyl viologen (1.0 mM) was studied. The formation of DCF in cyst cells was decreased with larger cell diameter. This decrease was also correlated with increases in astaxanthin content. Therefore, both cell diameter and the fluorescent DCF content of cyst cells would be good parameter to select astaxanthin-hyperproducing strains from native populations of H. pluvialis.     

Characterisation of anthocyanin–betalain mixtures for food colouring by chromatic and HPLC-DAD-MS analyses

Anthocyanins and betalains that are mutually exclusive in nature were mixed to investigate their potential synergism for food colouring purposes. The blends obtained from four commercial anthocyanic (black carrot, elderberry, sour cherry and strawberry) and one betalainic extract (red beet) were studied with respect to colour evolution over three weeks. While new colour shades were produced by blending anthocyanins with betalains, chroma and hue angle of the mixtures changed over time. The greatest number of new colour shades with acceptable chroma was obtained at pH 4.5 and 5, the stability maxima of red beet, succeeded by pH 3.0. Interestingly, at pH 3.5 only one combination yielded an acceptable chroma, whereas at pH 7 no mixture was stable. To determine individual anthocyanins and betalains in mixtures in a single run, a HPLC method was developed and combined with a mass spectrometer for the identification of specific pseudomolecular and daughter ions.     

Angiotensin-I converting enzyme inhibitory and antioxidant activities of protein hydrolysates from Phaseolus lunatus and Phaseolus vulgaris seeds

Phaseolus lunatus and Phaseolus vulgaris protein concentrates were hydrolyzed with the enzymes Alcalase^® and Flavourzyme^® at different reaction times, and the angiotensin-I converting enzyme (ACE-I) inhibitory activity, antioxidant properties and amino acid composition measured in the hydrolysates. With Alcalase^®, the highest degree of hydrolysis (DH) in P. lunatus was 37.94% at 45 min, and in P. vulgaris was 49.48% at 30 min. With Flavourzyme^®, the highest DH's were 22.03% and 26.05%, respectively, both at 90 min. ACE-I inhibitory activity in the Alcalase^® hydrolysates was IC₅₀ = 0.056 mg mL⁻¹ for P. lunatus at 90 min, and IC₅₀ = 0.061 mg mL⁻¹ for P. vulgaris at 60 min. In the Flavourzyme^® hydrolysates this activity was IC₅₀ = 0.0069 mg mL⁻¹ for P. lunatus at 90 min and IC₅₀ = 0.127 mg mL⁻¹ for P. vulgaris at 45 min. In SDS-PAGE, the hydrolysates exhibited low molecular weight bands. Antioxidant activity was 11.55 mmol L⁻¹ TEAC mg⁻¹ protein for P. lunatus with Flavourzyme^® at 90 min and 10.09 mmol L⁻¹ TEAC mg⁻¹ protein for P. vulgaris with Alcalase^® at 60 min. Amino acid composition exhibited high amino acid hydrophobic residues content.     

Potential use of a microalga (Chlorella vulgaris) in the pigmentation of rainbow trout (Oncorhynchus mykiss) muscle

The main purpose of this work was to evaluate the colouring effect of carotenoids present in dry biomass obtained from stressed cells ofChlorella vulgaris. Total carotenoids represented only 0.2% of dry algal biomass and consisted mainly of canthaxanthin, lutein, astaxanthin and their esters. — A rainbow trout feeding trial was conducted in order to investigate the effects of dietary algal incorporation as a pigment source, in comparison to that obtained with synthetic pigments, both astaxanthin and canthaxanthin. — The colour intensities measured in muscle were compared, in each case, with those which constitute the Salmonids Roche Colour Card, and proved similar after both 3 and 6 weeks feeding, with a significant increase detected, but at the end of experiment (9 weeks) the increase was shown only in spectrophotometric (quantitative) results.Chlorella vulgaris biomass, which is storable without any special precautions, thus appears to be a promising source of carotenoids to use in commercial fish finishing diets, quite comparable in efficiency with the existing synthetic pigments. Algal biomass micronization, which was also tried out, was not found to be advantageous for the efficiency of colouring, no evidence having been found of any difference in its absorption and in the distribution of pigment through muscle, even if a 150-g rainbow trout, which has a rather short digestive tract, was used.     

Association of genotype and preparation methods on the antioxidant activity, and antinutrients in common beans (Phaseolus vulgaris L.)

Three bean genotypes were selected in order to be evaluated, being IAPAR-81 (IAP), Uirapuru (UI) and BAF 55 (BAF). The samples were: raw (R), cooked without soaking (CWS), cooked with soaking water (CWSW) and cooked without soaking water (COSW). For the antioxidant activity, total phenolics, tannin and phytate analysis, the broths were separated from the grains and the soaking water separated from the COSW sample was analyzed. The CWSW and COSW grain samples showed higher antioxidant potential in the three genotypes (IAP: 0.037 and 0.039; UI: 0.035 and 0.040; BAF: 0.040 and 0.047, respectively). There were losses of total phenolics, tannin and phytate in all samples of the cooked beans demonstrating the importance of consumption and use of the cooked broths. The CWS was the preparation that preserved the majority of the nutrients in the grains, except for the antioxidant activity. In the broth, BAF 55 showed higher levels of tannin and phenolic compounds in all preparations. The analysis of the raw bean is important to know the nutritional value of the food, but the common bean is consumed cooked which modifies its characteristics as well as the availability of the nutrients to be absorbed.     

Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities

A total of 14 odoriferous angiospermic essential oils were tested against the toxigenic strain of Aspergillus flavus. The essential oil of Thymus vulgaris L. showed highest antifungal efficacy. The thyme oil absolutely inhibited the mycelial growth of A. flavus at 0.7 μl ml^− 1 and exhibited a broad fungitoxic spectrum against eight different food contaminating fungi viz. Fusarium oxysporum, Cladosporium herbarum, Curvularia lunata, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Alternaria alternata and Botryodiploidia theobromae. The oil also showed significant antiaflatoxigenic efficacy as it completely arrested the aflatoxin B₁ production at 0.6 μl ml^− 1. Thyme oil as fungitoxicant was also found superior over most of the prevalent synthetic fungicides. The LC₅₀ of thyme oil against mice was recorded as 7142.85 μl kg^− 1 body weight indicating its non-mammalian toxicity and strengthening its safe exploitation as preservative for stored food commodities. The findings recommend the thyme oil as potential botanical preservative in eco-friendly control of biodeterioration of food commodities during storage.

Industrial relevance

The thyme essential oil may be recommended for large scale application as a plant based preservative for stored food items because of its strong antifungal as well as antiaflatoxigenic efficacy. Because of broad antimicrobial spectrum, more efficacy over prevalent synthetic preservatives as well as non-mammalian toxicity, the thyme essential oil may be formulated as a safe and economical plant based preservative against post harvest fungal infestation and aflatoxin contamination of food commodities.     

Evaluation of the antioxidant activity of three microalgal species for use as dietary supplements and in the preservation of foods

The antioxidant activity of the microalgal ethanolic extracts of Porphyridium cruentum, Phaeodactylum tricornutum and Chlorella vulgaris was determined by means of the β-carotene–linoleate model system. The results show that the activity of C. vulgaris extract was higher than those obtained for the other microalgal extracts tested and for the synthetics BHA (butylated hydroxyanisole), and BHT (butylated hydroxytoluene). In addition, the major constituents present in the ethanolic extracts of the three microalgae species were analyzed by means of GC and GC–mass spectrometry. The results showed that the tested microalgae may be an important source of natural antioxidants, as an alternative to higher plants or the production by chemical synthesis.     

Effect of growth rate reduction and genetic modifications on acetate accumulation and biomass yields in Escherichia coli

Although acetate biosynthesis in Escherichia coli provides an important intermediary for ATP synthesis, its accumulation inhibits both cell growth and protein production. Since pyruvate provides the largest flux to acetate and is central to the problem of acetate production, acetate accumulation could be reduced or abolished if the pyruvate pool for the TCA cycle was reduced. To examine this possibility, various pyruvate kinase (pyk) and phosphotransferase system (pts) mutants were tested for acetate production in batch cultures with glucose as the only carbon source. The pykA pykF mutant exhibited significant reductions in the specific growth rate and acetate production compared with the wild-type strain. Interestingly, in the case of pts and pts pyk mutants in which increased biomass yields were observed in comparison with the wild-type strain, no acetate production was detected. Therefore, these mutants are potentially useful for higher production of recombinant proteins. The results from the continuous cultivation performed using the wild-type strain at various dilution rates, suggest acetate reduction as a consequence of both genetic changes and growth rate diminutions.     

Phytochemical composition and in vitro antimicrobial and antioxidant activities of some medicinal plants

Different parts of three plants (Primula auriculata, Fumaria vaillantii and Falcaria vulgaris) were extracted with three different solvents to yield 72 crude extracts. The phytochemical analysis (chemical screening, GC–MS) of three plants was investigated for their antioxidant and antibacterial activity using nine Gram-positive and Gram-negative bacteria. The principal antioxidant and antimicrobial components were determined using HPLC with UV detection. All extracts possessed antibacterial activity especially methanolic extracts from flowers of P. auriculata. The DPPH-radical scavenging assay exhibited high antioxidant activities in three plants (more than 80% at 50 μg). The F. vulgaris showed high content of carvacrol (29.8%) as main component. The contents of carvacrol and fumaric acid in the methanolic–water extracts were 1119 and 1966 mg/l respectively. Our results indicate that these plants would be able to promise sources of natural products with potential antibacterial and antioxidant activity.     

Antilisterial effects of free fatty acids and monolaurin in beef emulsions and hot dogs

This study was undertaken to screen fatty acids, conjugated isomers of linoleic acid (CLA), and monolaurin for antilisterial effects in broth, and to further test the active compounds in cooked comminuted beef and hot dogs. Capric, lauric, myristic, palmitic, stearic, oleic, linoleic, and linolenic acid, CLA and monolaurin were screened in sterile nutrient broth at concentrations of 5 to 700 μg/ml. The media were inoculated with 10³ cfu/ml of L. monocytogenes strain Scott A and incubated at 32°C for up to 8 days. Cell enumeration data showed that lauric acid was most inhibitory, followed by monolaurin, and capric acid. Tests in comminuted sterile beef stored at 5°C for 21 days showed log cfu/g of: 8.5 (control), 7.3 (500 μg/g lauric acid), and 4.7 (500 lauric acid+300 capric acid). Similar results were observed in beef hot dog emulsion to which lauricidin, lauric acid, capric acid, and the acid combination were added prior to heat treatment. At 500 μg/g, monolaurin and lauric acid caused similar delayed growth effects at 5°C, whereas the combination of the two acids showed enhanced inhibition on prolonged storage. Nonetheless, the observed 5-log increase in numbers of L. monocytogenes during 45 days of storage indicates limited control of the pathogen in refrigerated cooked meat products.     

Polyamines content in plant derived food: A comparison between soybean and Jerusalem artichoke

This paper aims to compare the dietary polyamine intake coming from foods derived from two different plants: soybean (Glycine max L.) well known and universally utilised both fresh and processed, and Jerusalem artichoke (Helianthus tuberosus L.) tuber, not yet well known and scarcely utilised in the everyday diet. Free, soluble and insoluble conjugated polyamines were determined in different soy-derived foodstuffs such as milk, tofu and fermented soy sauce, and in soybeans coming from two different organic experimental fields (Imola and Altedo, Bologna, Italy). Results show that free polyamines (in particular putrescine and spermidine) were present in relevant amounts especially in tofu and soy sauce. Conversely, the Helianthus parenchymatous medulla tissue, which is the only edible part of the tuber, contains very low levels of polyamines, which are instead preferentially accumulated in the buds. These data could suggest a preferential utilisation of Helianthus tuber in the diet of people with special needs, such as patients treated by chemotherapy and patients with diabetes.     

Impact of magnesium and mannose in the cultivation media on the magnesium biosorption, the biomass yield and on the cell wall structure of Candida utilis yeast

Binding capacity of Mg²⁺ ions, the biomass yield and the cell wall structure of Candida utilis ATCC 9950 cultivated in the media containing magnesium and mannose were evaluated in the study. Mannose has been added to two types of culture media: not enriched and enriched with Mg²⁺ ions. The YPD medium was used as a control while the experimental media (without or with Mg²⁺ ions) have been prepared by replacing glucose in the YPD medium by 1% (YPDM) or 2% (YPM) of mannose. The highest content of magnesium (5.39 and 5.42 mg/g_d.w.) as well as the highest biomass yield (15.22 and 14.03 g_d.w./L) were observed after 24 h of cultivation in the media enriched with magnesium and supplemented with mannose. The yeast cultivated for 48 h in those media were also characterized by thicker cell walls (74.3 and 67.7 nm). Introduction of mannose to the cultivation media was the factor that has influenced biosorption of magnesium ions as the result of twofold thickening of the mannoprotein layer of the cell wall of Candida utilis ATCC 9950 yeast.     

Compounds in functional food fenugreek spice exhibit anti-inflammatory and antioxidant activities

The seeds of fenugreek plant (Trigonella foenum-graecum) are widely used in the preparation of seasonings, pickles, curry powders and dietary supplements. The fenugreek seeds are also used in traditional medicine to relieve the common cold, arthritic pain and high blood sugar. Therefore, we have investigated the functional food quality of fenugreek seeds by determining the lipid peroxidation (LPO) and cyclooxyganase enzyme (COX) inhibitory activities of their hexane, ethyl acetate, methanolic and water extracts using MTT, LPO, COX-1 and -2 enzyme inhibitory assays. The extracts inhibited LPO by 55-95%, COX-1 by 6-87% and COX-2 by 36-70%, respectively, at 250 μg/ml. Bioassay-guided purification of these extracts yielded triglycerides (1-3), fatty acids (4-5), saccharides (6-8) and flavonoid-C-glycosides (9-11). The isolates, excluding the saccharides, inhibited LPO and COX-1 and -2 enzymes between the ranges of 8-89%, 4-51% and 15-70%, respectively, at 25 μg/ml. This is the first report of compounds 1-8 from fenugreek seeds and the biological activities described herein.     

Polyphenolic content and antioxidant activity of Eugenia pollicina leaf extract in vitro and in model emulsion systems

The polyphenolic profile of a leaf extract of the Mauritian endemic plant, Eugenia pollicina, was assessed as a source of natural antioxidants. The amounts of flavan-3-ol derivatives determined by HPLC, were in the order of (−)-epicatechin (EC) > (−)-epigallocatechin gallate (EGCG) > (+)-catechin (C) > (−)-epicatechin gallate (ECG) with the levels of Procyanidin B2 and B1 dimers ranging from 1 to 3 mg g⁻¹ FW. The trolox equivalent antioxidant capacity and ferric reducing antioxidant power values were 796 ??mol g⁻¹ FW and 302 ??mol g⁻¹ FW respectively. E. pollicina extracts also strongly inhibited the FeCl₃ and ascorbate-dependent microsome lipid peroxidation, a function that is linked to their flavonoid contents. The extent of DNA damage induced by the extract under study in the copper-phenanthroline assay was lower than the effect of a reference of 240 ??M ascorbate. E. pollicina extracts also inhibited lipid autoxidation in the 30% (v/v) olive oil and soybean oil oil-in-water (O/W) emulsions and was effective in slowing down the formation of hydroperoxides in the emulsions during 13 days storage at 40 °C as determined by the peroxide, conjugated diene and para-anisidine values. The high levels of total phenolics, flavonoids and procyanidins measured indicate that E. pollicina is a potential source of antioxidants relevant to the maintenance of oxidative stability of the food matrix, cosmetics and/or pharmaceutical preparations.