Carob Extract Research Articles

Ethanol production via repeated-batch fermentation from carob pod extract by using Saccharomyces cerevisiae in biofilm reactor

In present study, repeated-batch fermentation in biofilm reactor was evaluated for ethanol production by using carob extract. Response surface method (RSM) was used to determine optimum initial sugar concentration, pH, and agitation in the constructed biofilm reactor. The optimum conditions for ethanol production in biofilm reactor were determined as initial sugar content of 7.71°Bx, pH of 5.18 and agitation of 120rpm. The ethanol production (P), the yield (YP/S) and production rate (QP) were found as 24.51g/L 48.59% and 2.14g/L/h at the optimized conditions, respectively. The fermentation time for maximum ethanol production in carob extract was reduced to 12h by using a biofilm reactor compared the published data in the literature, which was 30 and 24h by using suspended and immobilized yeasts in a stirred tank reactor, respectively. Furthermore, the effect of various nitrogen sources and enrichment were also evaluated and YP/S, QS and QP were significantly decreased compared to the results obtained from optimum conditions in biofilm reactor. Overall, results showed that a biofilm reactor for ethanol production from carob extract can be successfully implemented in point of reducing fermentation time, eliminate re-inoculation of the typical batch fermentation as well as increasing production yield.

Gastroprotective effect of carob (Ceratonia siliqua L.) against ethanol-induced oxidative stress in rat.

BackgroundWe aimed in the present study, at investigating the gastroprotective effect of carob pods aqueous extract (CPAE) against ethanol-induced oxidative stress in rats as well as the mechanism implicated.MethodsAdult male wistar rats were used and divided into six groups of ten each: control, EtOH (80 % v/v, 4 g/kg b.w.), EtOH 80 % + various doses of CPAE (500, 1000 and 2000 mg/kg, b.w.) and EtOH + Famotidine (10 mg/kg, p.o.) Animals were perorally (p.o.) pre-treated with CPAE during 15 days and intoxicated with a single oral administration of EtOH (4 g/kg b.w.) for two hours.ResultsThe colorimetric analysis demonstrated that the CPAE exhibited an importance in vitro antioxidant activity against ABTS and DPPH radicals. We found that CPAE pretreatment in vivo, protected against EtOH-induced macroscopic and histological changes induced in stomach mucosa. Carob extract administration also protected against alcohol-induced volume gastric juice decrease. More importantly, We showed that CPAE counteracted EtOH-induced gastric lipoperoxidation, reversed the decrease of sulfhydryl groups (−SH) an hydrogen peroxide (H2O2) levels, and prevented the depletion of antioxidant enzyme activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx).ConclusionsThese findings suggest that CPAE exerted a potential gastro-protective effect against EtOH-induced oxidative stress in rats, due in part, to its antioxidants properties.

Chronic consumption of an inositol-enriched carob extract improves postprandial glycaemia and insulin sensitivity in healthy subjects: A randomized controlled trial

Inositols are thought to be mediators of the insulin signalling pathway. We assessed the effects of inositols on glycaemic control in fasting and postprandial states and evaluated lipoprotein profile and LDL particle size in healthy population. A 12-week double-blind clinical trial was performed with forty healthy subjects administered either an inositol-enriched beverage (IEB) -containing 2.23g of inositols in 250ml- or a sucrose-sweetened beverage (SB) twice a day. Anthropometric measurements, fasting glucose levels, insulin and HOMA-IR index, lipoprotein profile and postprandial glucose concentrations (measured using the continuous glucose monitoring system (CGMS)) were recorded throughout the intervention period. Following the 12-week trial subjects receiving the IEB exhibited a significant decrease in insulin, HOMA-IR and Apo B and an increase in LDL particle size, whereas the SB group showed increases in BMI and fasting glucose concentration. Analysis of postprandial glucose levels at breakfast, lunch and dinner revealed a mean reduction of glucose of ≈14% and a significant reduction in the area under the curve at 24h after consumption of the IEB. Our results show that chronic IEB supplementation induces a significant improvement in carbohydrated metabolism parameters in healthy subjects.

Antibacterial activity of carob (Ceratonia siliqua L.) extracts against phytopathogenic bacteria Pectobacterium atrosepticum

Acetone and ethanol extracts of carob (Ceratonia siliqua L.) leaf and pods were evaluated for their in vitro inhibitory ability against the pectinolytic Gram negative Pectobacterium atrosepticum (Pca, CFBP-5384) bacteria, the causal agent of potato soft rot. Potato (Solanum tuberosum, var nicola) tuber rot tissues obtained after 5 day bacterial inoculation was analyzed by LC–MS and GC–MS to study Pca pathogenicity. Trans/cis N-feruloylputrescine was identified in potato tuber after 5-day inoculation with Pca in a dark moist chamber. Although glycoalkoloid (α-chaconine and α-solanine) production increased due to Pca soft rot infection, it was not a resistance-determining factor. Many secondary metabolites were identified including the phytoalexins solavetivone and fatty acids responsible for plant defence responses. Acetone extract of carob leaf (FCA) exhibited the strongest inhibitory effect (IC50 = 1.5 mg/ml) and displayed synergistic antimicrobial effect in the presence of infected potato tuber extract (Pdt-Pca extract) against Pca. This synergy could be used in an integrated control program against potato soft rot pathogens, thereby reducing chemical treatments.

Extraction Kinetics of Phenolics from Carob (Ceratonia siliqua L.) Kibbles Using Environmentally Benign Solvents

A series of non-toxic and environmentally benign solvent systems, composed of water/ethanol, acidified with either acetic or citric acid, were tested for their efficiency in extracting polyphenols from dried carob kibbles. The best solvent system (30 % ethanol) was then used in a kinetic study, attempted by deploying non-linear regression. The leaching of polyphenols was found to obey 2nd-order kinetics, while the correlation of the total polyphenol yield at saturation with temperature obeyed a 3-parameter exponential growth model. The highest yield of total polyphenols at saturation was 53.76 mg gallic acid equivalents per gram of dry weight, achieved at 60 °C. The analysis of this extract using liquid chromatography–diode array–mass spectroscopy led to the tentative identification of gallic acid, myricetin 3-O-rhamnoside, quercetin 3-O-rhamnoside and quercetin p-hydroxy benzoyl 3-O-rhamnoside. The detection of the last compound in carob extracts is reported for the first time.

Effects on the lipid profile in humans of a polyphenol-rich carob (Ceratonia siliqua L.) extract in a dairy matrix like a functional food; a pilot study

The design of functional foods enriched in nutrients that favorably alter the lipid profile to prevent cardiovascular diseases and stimulate bowel function is of great interest. We have assayed a non-extractable-tannates-rich carob-fiber (PF-1®) in a milk matrix developed by Biosearch S.A. to discover its effects on the lipid profile and bowel function of human volunteers. A 4-week interventional study (400 mL daily consumption of this functional food, containing 20 g of PF-1®/L), was conducted: blood samples were analyzed for lipid profile, glucose, transaminases, creatinine and fat-soluble vitamins. The body-mass index and bowel function of the participants in the study were also measured. A tendency for triglyceride levels to diminish was observed in all participants (P = 0.066), and in the normal-cholesterol group in particular (P = 0.078). Another tendency to total cholesterol levels fell in the hypercholesterolemic group (P = 0.061) was also found. In the normal-cholesterol group, total cholesterol (CT), HDL-cholesterol and LDL-cholesterol levels significantly increased with the consumption of the functional food (P < 0.05). A better bowel function was also recorded by volunteers. This preliminary study highlights the possible positive influence of this functional food on the regulation of the lipid profile and bowel function in humans.

Ultrasound-assisted extraction of d-pinitol from carob pods using Response Surface Methodology

This study reports on the optimization of the conditions of extraction of d-pinitol compound from carob pods by using ultrasonication. For this purpose, the Box–Behnken design, which is a widely used form of Response Surface Methodology, was used to investigate the effects of parameters on the ultrasound-assisted extraction. Four independent variables, temperature (°C), ultrasonic power (W), dilution rate (material:water ratio) and time (min), were studied. The results showed that the highest d-pinitol concentration of 11.98g/L was obtained with an extraction temperature of 50°C, ultrasonic power of 207W, a dilution rate of 1:4 and an extraction time of 120min. It was found that the d-pinitol concentration of carob extract increased with the effect of ultrasonication during the extraction process. Therefore, ultrasound-assisted extraction could be used as an alternative to conventional hot water extraction with respect to the recovery of d-pinitol from carob pods with the advantages of lower extraction temperatures, shorter extraction time and reduced energy consumption.

Hepatoprotective effect of carob against acute ethanol-induced oxidative stress in rat

The present study was undertaken to determine whether subacute treatment with aqueous extract of carob (Ceratonia siliqua L.) pods (AECPs) protects against ethanol (EtOH)-induced oxidative stress in rat liver. Animals were divided into four groups: control, carob, EtOH and EtOH+carob. Wistar rats were intraperitoneally pretreated with AECP (600mg/kg body weight (bw)) during 7days and intoxicated for 6h by acute oral administration of EtOH (6g/kg bw) 24h after the last injection. We found that acute administration of EtOH leads to hepatotoxicity as monitored by the increase in the levels of hepatic marker aspartate aminotransferase and alanine aminotransferase as well as hepatic tissue injury. EtOH also increased the formation of malondialdehyde in the liver, indicating an increase in lipid peroxidation and depletion of antioxidant enzyme activities as superoxide dismutase, catalase and glutathione peroxidase. Subacute carob pretreatment prevented all the alterations induced by EtOH and returned their levels to near normal. Importantly, we showed that acute alcohol increased hepatic and plasmatic hydrogen peroxide and free iron levels. The carob pretreatment reversed EtOH effects to near control levels. These data suggest that carob could have a beneficial effect in inhibiting the oxidative damage induced by acute EtOH administration and that its mode of action may involve an opposite effect on plasma and tissue-free iron accumulation. Indeed, carob can be offered as a food additive to protect against EtOH-induced oxidative damage.

In vitro and in vivo antioxidant properties of Tunisian carob ( Ceratonia siliqua L.)

The present study aims at the quantification of phenolic compounds of Tunisian carob (Ceratonia siliqua L.) pods and the study of their in vitro and in vivo antioxidant activities in different extraction solvents. In this respect, we used 50 rats divided into five groups: Control and Carob-treated groups (100, 200, 600 and 1000 mg/kg, b.w.). Our results showed that the carob polar extracts are richer in total polyphenols, total flavonoids and condensed tannins than the nonpolar extracts with quantitative variation of phenolic compounds between seeds and pulp. In vitro, the determination of antioxidant capacity by 2,2'-azino-bis[3-ethylbenzthiazoline-6-sulphonic acid] (ABTS) method showed that carob extracts present high antioxidant potency in a solvent-compartment-dependent way compared to Trolox, an antioxidant reference molecule. In vivo, the subacute treatment within 7 days with aqueous extract of carob pods (AECP) decreased significantly and doses dependently the cerebral and myocardial lipid peroxidation as well as the level of hydrogen peroxide (H2O2) in kidney, liver and brain but not in heart. These data suggest that carob fruit presents an in vitro an in vivo antioxidant effects and might be proposed as a food additive to protect against oxidative stress damage. Key words: Carob, phenolic compounds, antioxidant properties, rat.

Inhibitory Effect of Carob (Ceratonia siliqua) Leaves Methanolic Extract on Listeria monocytogenes

In recent years, there has been great development in the search for new natural compounds for food preservation aimed at a partial or total replacement of currently popular antimicrobial chemicals. Carob (Ceratonia siliqua) offers a natural promising alternative for food safety and bioconservation. In this work, the methanolic extract of carob leaves (MECL) was tested for the ability to inhibit the growth of a range of microorganisms. MECL inhibited the growth of Listeria monocytogenes at 28.12 μg/mL by the broth microdilution method. The effect of this bacteriostatic concentration on the growth of this bacterium revealed a pattern of inhibition characterized by (a) a resumed growth phase, which showed a lower rate of growth if compared with controls; and (b) first a lag and then a stationary phase at a lower bacterium concentration. The study of the chemical composition of MECL by high-performance liquid chromatography and liquid chromatography/mass spectrometry showed the presence of gallic acid, (-)-epigallocatechin-3-gallate, myricitrin, isoquercitin, catechin, chlorogenic acid, and malic acid. L. monocytogenes growth inhibition was recorded for myricitrin and gallic acid at 450 μg/mL and for (-)-epigallocatechin-3-gallate and isoquercitin, respectively, at 225 and 112.5 μg/mL. Taking into account that proline is a ligand of proline dehydrogenase (PDH), the use of this compound leads us to hypothesize the mode of action of MECL constituents.

Potential uses of microorganisms for d-pinitol purification from carob extracts

Potential uses of microorganisms for d-pinitol purification from carob extracts

Optimization of ethanol production from carob pod extract using immobilized Saccharomyces cerevisiae cells in a stirred tank bioreactor

In this study, optimization of ethanol production from carob pod extract was carried out by immobilized Saccharomyces cerevisiae. Results showed that Ca-alginate concentration and the amount of immobilized cells had significant effects on yield. Optimum conditions for ethanol fermentation were determined to be 2% Ca-alginate concentration, 150rpm agitation rate, 5% yeast cells entrapped in beads and pH 5.5. After validation experiments; ethanol concentration, yield, production rate and sugar utilization rate were respectively 40.10g/L, 46.32%, 3.19g/L/h and 90.66%; and the fermentation time was decreased to 24h. In addition, the immobilized cells were shown to be reusable for five cycles, though a decrease in yield was observed. Finally, carob pod extract was used for ethanol fermentation by controlled and uncontrolled pH without any enrichment, and the results suggest that carob extract can be utilized effectively by immobilized-cell fermentation without the use of enrichments to facilitate yeast growth.

Kinetics of sugars consumption and ethanol inhibition in carob pulp fermentation by Saccharomyces cerevisiae in batch and fed-batch cultures

The waste materials from the carob processing industry are a potential resource for second-generation bioethanol production. These by-products are small carob kibbles with a high content of soluble sugars (45-50%). Batch and fed-batch Saccharomyces cerevisiae fermentations of high density sugar from carob pods were analyzed in terms of the kinetics of sugars consumption and ethanol inhibition. In all the batch runs, 90-95% of the total sugar was consumed and transformed into ethanol with a yield close to the theoretical maximum (0.47-0.50g/g), and a final ethanol concentration of 100-110g/l. In fed-batch runs, fresh carob extract was added when glucose had been consumed. This addition and the subsequent decrease of ethanol concentrations by dilution increased the final ethanol production up to 130g/l. It seems that invertase activity and yeast tolerance to ethanol are the main factors to be controlled in carob fermentations. The efficiency of highly concentrated carob fermentation makes it a very promising process for use in a second-generation ethanol biorefinery.

Enhanced Lactic Acid Production from Carob Extract by Lactobacillus casei Using Invertase Pretreatment

Carob has been widely grown in the Mediterranean region for centuries. It has been regarded as only a forest tree and has been neglected for other economical benefits. However, in recent years, it has gained attention for several applications. In this study, carob extract has been used as carbon source in fermentation. Lactic acid is considered to be an industrial chemical used in food, textile and pharmaceutical industries. Lactic acid demand has recently increased because of its role as a monomer in the production of biodegradable polylactic acid (PLA), which is known as a sustainable and degradable bioplastic material. Therefore, this study was undertaken to evaluate the potential of carob extract for lactic acid production. Based on our previous study, the optimum extraction conditions were used to obtain the carob extract with high sugar content. Then, obtained extract was utilized by Lactobacillus casei for production of lactic acid by using suspended-cell fermentation. Although carob pod contains 40–50% sugars (especially, sucrose), L. casei could not metabolize them in this complex form. Therefore, sucrose in extract was converted into the monosaccharides by using invertase enzyme prior to fermentation. Also, in order to determine economical appropriate nitrogen sources, four different nitrogen sources were evaluated. The maximum lactic acid concentration and yield for carob extract with yeast extract was 31.35 (g/L) and 68.79% compared with 59.27 (g/L) and 66.70% after using invertase enzyme. Results clearly showed that carob pods can be a good feedstock for lactic acid production by L. casei.

Carob pulp as raw material for production of the biocontrol agent P. agglomerans PBC-1

Large-scale production has been the major obstacle to the success of many biopesticides. The spreading of microbial biocontrol agents against postharvest disease, as a safe and environmentally friendly alternative to synthetic fungicides, is quite dependent on their industrial mass production from low-cost raw materials. Considerable interest has been shown in using agricultural waste products and by-products from food industry as nitrogen and carbon sources. In this work, carob pulp aqueous extracts were used as carbon source in the production of the biocontrol agent Pantoea agglomerans PBC-1. Optimal sugar extraction was achieved at a solid/liquid ratio of 1:10 (w/v), at 25°C, for 1 h. Batch experiments were performed in shake flasks, at different concentrations and in stirred reactors at two initial inoculums concentrations, 10(6) and 10(7) cfu ml(-1). The initial sugar concentration of 5 g l(-1) allowed rapid growth (0.16 h(-1)) and high biomass productivity (0.28 g l(-1) h(-1)) and was chosen as the value for use in stirred reactor experiments. After 22 and 32 h of fermentation the viable population reached was 3.2 × 10(9) and 6.2 × 10(9) cfu ml(-1) in the fermenter inoculated at 10(6) cfu ml(-1) and 2.7 × 10(9) and 6.7 × 10(9) cfu ml(-1) in the bioreactor inoculated at 10(7) cfu ml(-1). A 78% reduction of the pathogen incidence was achieved with PBC-1 at 1 × 10(8) cfu ml(-1), grown in medium with carob extracts, on artificially wounded apples stored after 7 days at 25°C against P. expansum.

Promising antioxidant and cytotoxic activities of the aqueous ethanolic extract of carob leaves

Plants are endless reservoir for several phytochemicals including antioxidant and cytotoxic drugs. The objective of this study was the investigation of the antioxidant and the cytotoxic activities (against Vero and HEp-2 cell lines) of the aqueous ethanolic extract of carob leaves. The extract inhibited xanthine oxidase at IC50 of 244 μg/ml. There is a marked cytotoxic activity of crude aqueous extract of carob on both mammalian cell lines (Vero and HEp-2) and the effect is more obvious on the human cell line (HEp-2). The results revealed that the crude aqueous extract of carob leaves could be a promising source for antioxidant as well as antitumor bioactive agents. Key words: Carob leaves, aqueous ethanolic extract, Ceratonia siliqua, antioxidant, xanthine oxidase, cytotoxicy.

Ethanol production from carob extract by using Saccharomyces cerevisiae

Carob has been widely grown in the Mediterranean region for a long time. It has been regarded as only a forest tree and has been neglected for other economical benefits. However, in recent years, this fruit has gained attention for several applications. As petroleum has become depleted, renewable energy production has started to gain attention all over the world; including the production of ethanol from underutilized agricultural products such as carob. In this project, the optimum extraction conditions were determined for the carob fruit by using the response surface design method. The obtained extract was utilized for production of ethanol by using suspended Saccharomyces cerevisiae fermentation. The effect of various fermentation parameters such as pH, media content and inoculum size were evaluated for ethanol fermentation in carob extract. Also, in order to determine economically appropriate nitrogen sources, four different nitrogen sources were evaluated. The optimum extraction condition for carob extract was determined to be 80 °C, 2 h in 1:4 dilution rate (fruit: water ratio) according to the result of response surface analysis (115.3 g/L). When the fermentation with pH at 5.5 was applied, the final ethanol concentration and production rates were 42.6 g/L and 3.37 g/L/h, respectively, which were higher than using an uncontrolled pH. Among inoculum sizes of 1%, 3%, and 5%, 3% was determined as the best inoculum size. The maximum production rate and final ethanol concentration were 3.48 g/L/h and 44.51%, respectively, with an alternative nitrogen source of meat–bone meal. Overall, this study suggested that carob extract can be utilized for production of ethanol in order to meet the demands of renewable energy.

중탄산소듐 용액에서 납의 부식방지제인 자연산물질들

0.1 M 중탄산소듐 용액에서 납전극부식에 대한 로소니아, 감초뿌리 및 카로브 추출물수용액의 부식방지작용에 대한것을 정전류극성방법을 사용해 연구하였다. 이들 물질들의 부식방지작용을 지적해주는 것으로 이들 물질들이 존재할 때 부식속도가 감소하는 것이 발견되었다. 부식방지효율은 추출물농도가 증가하면 증가한다. 이들 추출물의 부식방지작용은 납표면에 이들 추출물이 흡착되어 물질 및 하전이동의 장벽을 만들어 나타내는 것으로 설명되었다. 이들 추출물의 납표면에 흡착은 자발적으로 일어나며, Freundlich 등온흡착을 따르는 것으로 발견되었다. 또한, 변전위양극분극법을 사용해 이들 추출물은 염소를 포함한 용액에서 납의 움푹패이게 하는 부식으로부터 잘 보호해주는 것이 발견되었다. The inhibitive action of the aqueous extract of lawsonia, licorice root and carob toward the corrosion of tin electrode in 0.1 M <TEX>$NaHCO_3$</TEX> solutions was investigated using galvanostatic polarization measurements. It was found that the corrosion rate decreases in the presence of these extracts indicating the inhibiting of these compounds. The inhibition efficiency increases with increasing extract concentration. The inhibition action of these extracts was explained in view of adsorption of its compounds onto the tin surface, making a barrier to mass and charge transfer. The adsorption of these extracts on the tin surface was found to be a spontaneous process and follow Freundlich adsorption isotherm. It is also found that these extracts provide a good protection to tin against pitting corrosion in chloride containing solution using potentiodynamic anodic polarization technique.

New insight into the influence of carob extract and gallic acid on hemin induced modulation of HT29 cell growth parameters

Red meat intake is associated with an increased risk of developing cancer. This is possibly related to the heme content of red meat. Plant derived polyphenols might protect from cancer development via their antioxidant activities. In this study, the impact of an aqueous extract of carob (CE) on hemin-modulated proliferation was investigated. CE, gallic acid (GA) and a known iron chelator (deferoxamine: DFO) significantly reduced the number of human colon cancer HT29 cells. CE and GA were more effective under serum-free conditions than in normal cell culture medium. These effects were abolished by addition of 1 μM hemin at low concentrations of CE and GA. At higher concentrations of CE and GA, both substances reduced cell number despite hemin supplementation. Effects of CE, GA and DFO on cell number could not be linked to iron chelation even though CE and DFO were capable of chelating iron. Furthermore, the effects of high CE concentration point to antioxidative effects other than iron chelation. However, a connection to a reduction of colorectal cancer risk due to consumption of meat with high heme content by CE could not be drawn, since the effective concentrations are beyond the physiologically relevant concentrations.

Does an Extract of Carob (Ceratonia siliqua L.) Have Chemopreventive Potential Related To Oxidative Stress and Drug Metabolism in Human Colon Cells?

Phenolic ingredients of an aqueous carob extract are well characterized and consist of mainly gallic acid (GA). In order to assess possible chemopreventive mechanisms of carob, which can be used as a cacao substitute, effects on expression of genes related to stress response and drug metabolism were studied using human colon cell lines of different transformation state (LT97 and HT29). Stress-related genes, namely catalase (CAT) and superoxide dismutase (SOD2), were induced by carob extract and GA in LT97 adenoma, but not in HT29 carcinoma cells. Although corresponding protein products and enzyme activities were not elevated, pretreatment with carob extract and GA for 24 h reduced DNA damage in cells challenged with hydrogen peroxide (H(2)O(2)). In conclusion, carob extract and its major phenolic ingredient GA modulate gene expression and protect colon adenoma cells from genotoxic impact of H(2)O(2). Upregulation of stress-response genes could not be related to functional consequences.