Carob Pod Extract Research Articles

Immature carob pods extract and its fractions prevent lipid metabolism disorders and lipoprotein-rich plasma oxidation in mice: A phytochemical and pharmacological study

Ethnopharmacological relevance: In Morocco carob fruits are used traditionally to treat hypercholesterolemia, diabetes and related diseases. Aims: This study was designed to evaluate the hypolipidemic activity of Ceratonia siliqua green pods extract and its fractions in Triton WR-1339 and high fat/cholesterol diet (HFCD) induced hyperlipidemia mice, as well as their ability to prevent lipoproteins oxidation in vitro. Materials and methods: High performance liquid chromatography (HPLC) analysis was used to determine the phenolic composition of the immature carob pods extract (HWCE). Antioxidant activities were evaluated using the DPPH radical scavenging test as well as MDA measurement in oxidized lipoprotein rich plasma. Plasma lipids, glucose and biliary total cholesterol, as well as lipids level in liver and feces, were analyzed. The acute oral toxicity was performed in mice single dosed with the HWCE at 2000 and 5000 mg/kg body weight. Results: HPLC analysis shows that gallic acid is the main phenolic compound in the HWCE. The acute oral toxicity assessment revealed that the HWCE is not toxic (LD50 is greater than 5000 mg/kg body weight). In the acute hypolipidemic study, mice treated with the HWCE and its fractions exhibited a significant (P < 0.001) reduction in plasma total cholesterol (TC), triglycerides (TG) and low density lipoprotein-cholesterol (LDL-C) levels. Importantly, immature carob aqueous extract was more effective in lowering mice hypercholesterolemia than its fractions. Indeed, mice fed the HFCD for 12 weeks showed a significant raise in plasma TC, TG and LDL-C, as well as in hepatic and fecal TC and TG levels. The HWCE at 100 and 200 mg/kg body weight significantly (P < 0.001) reversed the plasmatic levels of these lipid parameters, increased plasma HDL-C level, reduced hepatic lipids accumulation, but increased cholesterol level in the bile and fecal lipids excretion. The HWCE decreased also the atherogenic index, the LDL-C/HDL-C ratio and plasma glucose level after 12 weeks’ experiment. On the other hand, the HWCE was more effective in preventing mice lipoprotein-rich plasma oxidation than its fractions, with a concentration-dependent manner. Conclusion: C. siliqua green fruits extract could be effective in preventing atherosclerosis and related cardiovascular complications through the inhibition of lipoprotein oxidation and cholesterol clearance.

Protective Effect of Carob Pods Extract on Esomeprazole-Induced Changes on the Renal Cortex of Rats: Histological, Immunohistochemical and Statistical Study

Protective Effect of Carob Pods Extract on Esomeprazole-Induced Changes on the Renal Cortex of Rats: Histological, Immunohistochemical and Statistical Study

Optimization and kinetic modeling of media composition for hyaluronic acid production from carob extract with Streptococcus zooepidemicus.

Hyaluronic acid (HA), a mucopolysaccharide belonging to the glycosaminoglycan family, consists of repeating disaccharide units and has been used directly or indirectly in numerous human health practices. This study focused on evaluating carob pods for microbial HA production and kinetic modeling of HA fermentation. Therefore, the optimal medium composition was determined using Plackett-Burman Design (PBD) for HA production from carob extract with Streptococcus zooepidemicus. Maximum HA production of shake flask fermentation was 2.6g/L (1.25 × 106) in the optimum medium, comprising 10°Bx of carob pods extract, 0.5g/L of MgSO4.7H2O, 10.0g/L of casein, 2.5g/L of KH2PO4, 2.0g/L of NaCl, 1.5g/L of K2HPO4, 0.002g/L of FeSO4 and 10.0g/L of beef extract. In the continuation of the study, the fermentation performed with the optimal medium composition was modeled using three different models including the logistic model for biomass production, the Luedeking-Piret model for HA production, and the modified Luedeking-Piret model for substrate consumption. Based on the results, the experimental HA production data agreed with the Luedeking-Piret model with an R2 of 0.989. Since the α value was 63-fold higher than the value of β, the HA production is growth-associated. Consequently, carob extract can be evaluated as a promising carbon source for producing HA.

A novel rotating wide gap annular bioreactor (Taylor-Couette type flow) for polyhydroxybutyrate production by Ralstonia eutropha using carob pod extract

An annular bioreactor (ABR) with wide gap was used for PHB production from Ralstonia eutropha. Hydrodynamic studies demonstrated the uniform distribution of fluid in the ABR due to the Taylor-Couette flow. Thereafter, the ABR was operated at different agitation and sparging rates to study its effect on R. eutropha growth and PHB production. The ABR operated at 500 rpm with air sparge rate of 0.8 vvm yielded a maximum PHB concentration of 14.89 g/L, which was nearly 1.4 times that obtained using a conventional stirred-tank bioreactor (STBR). Furthermore, performances of the bioreactors were compared by operating the reactors under fed-batch mode. At the end of 90 h of operation, the ABR resulted in a very high PHB production of 70.8 g/L. But STBR resulted in a low PHB concentration of 44.2 g/L. The superior performance was due to enhanced oxygen and nutrient mass transfer in the ABR.

Techno-economic assessment of a sustainable and cost-effective bioprocess for large scale production of polyhydroxybutyrate

The rapid depletion of crude-oil resource which sustains a conventional petroleum refinery together with its environmental impact has led to the search for more sustainable alternatives. In this context, biorefinery serves to fulfil the aim by utilizing waste resources. Hence, this study focused on techno-economic assessment of PHB production at large scale from waste carob pods in a closed-loop biorefinery setup. Firstly, the use of pure sugars in SC1 was shifted to use of carob pods as feedstock in SC2, upgradation of stirred tank bioreactor with novel annular gap bioreactor in SC3 and replacing the conventional centrifugation process with the upcoming ceramic membrane separation process in SC4. An Aspen plus™ flowsheet was developed by including the aforementioned novel strategies for PHB production. The effectiveness of PHB production under various scenarios was evaluated based on its pay-out period and turnover accumulated at the end of 7th year of a PHB plant operation. Instead of pure sugars as the feedstock (SC1), carob pod extract (SC2) reduced the pay-out period from 12.6 to 6.8 years. Likewise, switching onto ABR from the conventional STBR further decreased the pay-out period to 4.8 years. Finally, the use of ceramic membranes (SC4) instead of centrifugation resulted in a similar pay-out period of 4.8 years with increased turnover of about 1.4 billion USD. Thus, the use of carob pods along with an improved PHB titre in ABR and incorporation of affordable ceramic membrane technology for PHB rich biomass separation resulted in a highly cost-effective PHB production strategy.

A closed-loop biorefinery approach for polyhydroxybutyrate (PHB) production using sugars from carob pods as the sole raw material and downstream processing using the co-product lignin

A novel closed-loop biorefinery model using carob pods as the feed material was developed for PHB production. The carob pods were delignified, and as the second step, sugars present in the delignified carob pods were extracted using water. Ralstonia eutropha and Bacillus megaterium were cultivated on the carob pod extract and its performance was evaluated using Taguchi experimental design. R. eutropha outperformed the B. megaterium in terms of its capability to grow at a maximum initial sugar concentration of 40 g L−1 with a maximum PHB production of 12.2 g L−1. Finally, the concentrated lignin from the first step was diluted with different proportion of chloroform to extract PHB from the bacterial biomass. The PHB yield and purity obtained were more than 90% respectively using either R. eutropha or B. megaterium. Properties of the PHB produced in this study were examined to establish its application potential.

FARKLI HİDROLİK ALIKONMA SÜRELERİNDE KEÇİBOYNUZU EKSTRAKTI BESİYERİNDE SÜREKLİ ETANOL FERMANTASYONU

Production of bioethanol is one of the important bioprocesses for the energy industry to provide inexpensive renewable resources all over the world. In this context, this research was organized for continuous ethanol fermentation from carob pod extract which is an inexpensive carbon source by free or immobilized S. cerevisiae cells. Continuous ethanol fermentations were performed with different HRT (from 4 to 20 h) and optimal HRT were 8 h for the free cell, and 6.67 h for immobilized cell, respectively. The highest volumetric ethanol productivities for free cell and immobilized cell fermentations were 3.12 g/L/h and 3.37 g/L/h at HRT of 5.71 h, respectively. All kinetic parameters clearly showed that both cell types can be used for ethanol fermentation, and immobilized S. cerevisiae ethanol fermentation can be operated at higher dilution rates independent of biomass than a free cell.

Using agro-industrial wastes for mixotrophic growth and lipids production by the green microalga Chlorella sorokiniana

There has been growing interest in the use of microalgae for the production of biofuels, but production costs continue to be too high to compete with fossil fuel prices. One of the main limitations for photobioreactor productivity is light shielding, especially at high cell densities. The growth of the green microalga Chlorella sorokiniana, a robust industrial species, has been evaluated under different trophic conditions with traditional carbon sources, such as glucose and sucrose, and alternative low cost carbon sources, such as carob pod extract, industrial glycerol and acetate-rich oxidized wine waste lees. The mixotrophic cultivation of this microalga with wine waste lees alleviated the problems of light shielding observed in photoautotrophic cultures, improving specific growth rate (0.052 h-1) compared with the other organic sources. The fed-batch mixotrophic culture of Chlorella sorokiniana in a 2 L stirred tank reactor, with optimized nutritional conditions, 100 mM of acetate coming from the oxidized wine waste lees and 30 mM of ammonium, produced an algal biomass concentration of 11 g L-1 with a lipid content of 38 % (w/w). This fed-batch strategy has been found to be a very effective means to enhance the biomass and neutral lipid productivity.

English

English

Carob (Ceratonia siliqua L.) Prevents Short-Term Memory Deficit Induced by Chronic Stress in Rats.

Long-term exposure to stressful conditions could impair the normal brain structure and function, specifically the hippocampus-dependent memory. This impairment could be attributed to a decrease in brain-derived neurotrophic factor (BDNF) levels during chronic stress. Knowing that carob [Ceratonia siliqua L. (Fabaceae)] is rich in a wide variety of polyphenols with a high antioxidant value, we hypothesized that the methanolic carob extract (C. siliqua) pods will prevent stress-induced memory impairment. Hence, the methanolic extract of carob pods was investigated for its ability to enhance learning and memory as well as to protect from memory impairment in normal stressed animals. Rats were chronically stressed for 7weeks via the intruder stress model. Carob extract was administered to animals via intraperitoneal (i.p.) route at a daily dose of 50mg/kg. Radial arm water maze (RAWM) was utilized to test for spatial learning and memory. In addition, brain tissues were dissected to determine BDNF levels. Chronic stress (CS) impaired short-term spatial memory (number of committed errors: P < 0.05, days to criterion (DTC): P < 0.001). Animal treatment with carob pod extract prevented the short-term memory impairment induced by CS (P < 0.05), while such treatment showed no effect on memory functions of unstressed rats. Moreover, carob pod extract prevented the reduction in the hippocampal BDNF (P < 0.05) induced by chronic stress exposure. In conclusion, CS impaired short-term memory function, while methanolic extract of carob pods prevented this impairment, probably as a result of preventing reduction in BDNF levels in the hippocampus.

Carob as a carbon source for fermentation technology

Abstract Carob (Ceratonia siliqua L.), have wild and cultivated types, is an evergreen tree and widely grown in Mediterranean countries. Wild or cultivated carob has a high total dry weight content (approximately 91–92%) which primarily consist of sucrose, glucose, fructose, and also incorporate minor amounts of minerals, amino acids, and phenolic compounds. The seeds are generally used for the production of locust bean gum and the kibble is used to produce carob powder in western countries or “pekmez” in Turkey. Finding new carbon and nitrogen sources is important for lab-scale research to provide cheaper substrates for biotechnological processes to reduce costs. In this sense, carob is one of the important sources for any kind of fermentation processes with its rich nutritional ingredients. Carob pod extracts have been used in many lab-scale studies to produce ethanol, citric acid, lactic acid, mannanase, microbial cell protein, and other value-added products by yeasts, algae or fungi in recent years. In this review, we tried that organize all researches about carob pod extract fermentation to prove the suitability of carob pod for biotechnological processes.

Performance comparison of precipitation strategies for recovering succinic acid from carob pod-based fermentation broths

ABSTRACTExperiments were performed for comparing and selecting the most appropriate precipitation strategy for succinic acid (SA) recovery from carob pod extract fermentation broths. The performances of three downstream options – employing calcium hydroxide, magnesium hydroxide or ammonia – were investigated and compared from operational, techno-economical and sustainability viewpoints. The highest SA recovery (84.3%) was obtained with ammonia, whereas the calcium-based treatment proved inadequate. Sustainability indicators favoured magnesium-based treatment, closely followed by the ammonia-based one. A preliminary economic analysis favours magnesium-based treatment, which appears to be the most feasible option. However, materials recycling and possible by-products commercialization could well position also ammonia-based treatment.

Hepatoprotective Effect of Carob Pods Extract (Ceratonia siliqua L.) against Cyclophosphamide Induced Alterations in Rats

Drugs used for treatment of cancerous diseases by mean of chemotherapy are often limited due to their severe undesirable side effects in multiple organs. Cyclophosphamide (CYP) belongs to class of the cytotoxic bifunctional alkylating agents. In the current study, it was revealed that CYP caused significant (P˂0.05) elevation in levels of different biochemical measurements. Carob extract exhibited beneficial effect through lowering of all elevated parameters. Furthermore, CYP caused decline in total antioxidant capacity (TAC) level in association with elevation of lipid peroxidation (LPO) level and significantly (P˂0.05) in liver tissue. Carob extract restored TAC and lowered LPO level. CYP caused several histopathological alterations in the hepatocytes and carob extract minimized severity of these alterations. The similarity index percent (SI %) in the native electrophoretic protein, lipoprotein and calcium moiety of protein patterns was represented by lowest values (87.50, 66.67 and 70.59, respectively) with the CYP-treated group. The SI % values increased in all carob treated groups. In all electrophoretic isoenzymes, the SI % value was represented by the lowest value (60.00, 66.67, 66.67 and 53.33 respectively with catalase (CAT), peroxidase (GPx), α- and β-esterase (EST) patterns) in CYP-treated group. While in all carob treated groups, the SI % value reached the highest value (100.00). Furthermore, CYP induced cleavage of the genomic DNA and the carob extract maintained the DNA integrity. The study concluded that carob showed ameliorative effect against alterations induced by CYP at biochemical, histopathological and molecular levels in liver tissue of rats.

Ameliorative Effect of Carob Pods Extract (Ceratonia siliqua L.) against Cyclophosphamide Induced Alterations in Bone Marrow and Spleen of Rats

Many anticancer drugs including cyclophosphamide (CYP) exhibit potentially life-threatening effects on the immune and hematopoietic systems. The spleen and lymphatic tissues are the most sensitive components of immune system. In the present study, it was verified that CYP caused significant (P

Enhanced β-mannanase production from alternative sources by recombinant Aspergillus sojae

β-mannanases can degrade galactomannans to mannose and it has been used in various application areas. The aim of this study was to produce the β-mannanase from carob pod extract including different nitrogen sources. The best operation values for fermentation were determined to be 8% initial sugar concentration with 0.5% yeast extract, 100 r.p.m., and 7% inoculation rate, which yielded the maximum β-mannanase activity as 423.60 U ml–1. Effects of nitrogen sources on β-mannanase activity were also studied and it reached 695.6 U ml−1 by using 0.5% of ammonium nitrate as the nitrogen source at the determined optimum conditions. Results also showed that meat bone meal and soybean meal could be used as cost effective nitrogen sources based on achieved β-mannanase activity.

Controlling filamentous fungi morphology with microparticles to enhanced β-mannanase production.

β-mannanase was produced mainly by Aspergillus species and can degrade the β-1,4-mannose linkages of galactomannans. This study was undertaken to enhance mannanase production using talcum and aluminum oxide as the microparticles, which control cell morphology of recombinant Aspergillus sojae in glucose and carob extract medium. Both microparticles improved fungal growth in glucose and carob pod extract medium. Aluminum oxide (1g/L) was the best agent for glucose medium which resulted in 514.0U/ml. However, the highest mannanase activity was found as 568.7U/ml with 5g/L of talcum in carob extract medium. Increase in microparticle concentration resulted in decreasing the pellet size diameter. Furthermore, more than 10g/L of talcum addition changed the filamentous fungi growth type from pellet to pellet/mycelium mixture. Results showed that right type and concentration of microparticle in fermentation media improved the mannanase activity and production rate by controlling the growth morphology.

Membrane assisted recovery and purification of bio-based succinic acid for improved process sustainability

Three membrane-assisted technologies – electrodialysis, nanofiltration, and Donnan dialysis – were investigated with the objective of developing a green and sustainable integrated downstream process for the recovery of biologically produced succinic acid from carob pod-based fermentation broths. For electrodialysis, heterogeneous and homogenous anion-exchange membranes (including such with monovalent-anion-permselective transport properties) were tested with synthetic model solutions in order to select the most suitable membrane. For nanofiltration, six polymeric membranes were tested to filter the obtained electrodialysis concentrates, and according to the results obtained, three of them (NF270, NF-DK and NF-DL) were selected for further experiments. The performance of these membranes was compared in a diafiltration operation mode with model carboxylates solutions and with simulated fermentation broths, all showing rejections above 90% for succinate and reaching negative rejections for acetate and formate. However, reduced fluxes and fouling were observed in the experiment with simulated fermentation broth containing a carob pod extract. Therefore this medium was treated by electrodialysis, producing a concentrate rich in ionic species, including succinate, but leaving behind the non-ionic and high-molecular mass species. This concentrate was treated by nanofiltration, thus achieving an almost complete removal of accompanying formate and acetate species. Finally, Donnan dialysis was employed to replace the metallic cations in the succinate salts in the nanofiltration retentate by H+. The results showed approximately 76% of cations removal, which was traduced in the same percentage of obtained free succinic acid. Thus, the proposed integrated membrane-assisted process represents a promising new alternative to the existing downstream technologies for bio-based succinic acid recovery.

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.

Fermentative Hydrogen Production from Carob Pod: A Typical Mediterranean Forest Fruit

The carob-tree (Ceratonica siliqua L.) grows in arid to semi-arid Mediterranean areas and adapts well to poor soils. Carob pod, the fruit of carob tree is a sugar-rich biomass which may theoretically be ideal for biofuel production. During this study, the potential of hydrogen production from carob pod was evaluated in an anaerobic continuous stirred tank bioreactor using mixed microflora. A carob pod extract consisting of soluble carbohydrates at a concentration of 42.6 g/L was obtained from a standard extraction process. The reactor was operated at a hydraulic retention time (HRT) of 36 and 18 h and an organic loading rate of 155.8 and 311.6 mmol glucose/L/days respectively. The results showed a maximum hydrogen production yield of 8.4 L H2 per kg of carob pod biomass or 0.43 mmol H2 per mmol glucose consumed at the HRT of 18 h. Butyric acid was found to be the dominant metabolic product at both examined HRTs while propionic acid production decreased as HRT decreased. The present work suggests that carob tree, an alternative dryland forest crop, can be utilized for the efficient production of biohydrogen.

CitricAcid Production from Carob Pod Extract by Aspergillus. Niger

The present study was undertaken to study the production of citric acid by the fungus Aspergillus niger using carob pod extract as a carbon source for fungal growth and production of citric acid. High yields of citric acid were achieved after six days of incubation when 10% of carob pod extract as a carbon source present in fermentation media with pH value of 3.16 and the amount of yield being12.25g\l, (55.48%). The results also revealed that the accumulation of citric acid by the fungus was affected by the source and concentration of nitrogen, as well as high production of the acid was obtained in fermentation medium containing ammonium sulphate, in which the accumulated amount increased up to 18.25g\l, (70.16%). Moreover, the addition of 0.05% calcium chloride and 2% ethanol to the fermentation medium highly stimulated citric acid accumulation by the fungus to reach 21.06g\l and this amount equivalent to 113.22% of the biomass dry weight.