Triterpenic acids in table olives

An experimental investigation was carried out for the first time on the triterpenic acids in table olives. Maslinic acid was found in a higher concentration than oleanolic acid in the flesh of 17, unprocessed olive varieties, with the Picual and the Manzanilla varieties showing the highest and almost the lowest contents, respectively. The level of triterpenic acids in several types of commercial black and green olives ranged from 460 to 1470 mg/kg fruit, which represents a much higher value than reported for virgin olive oils. In fact, the NaOH treatment employed to debitter black and green olives reduced the concentrations of these substances in the flesh because of their solubilisation into alkaline solutions. Thus, natural black olives, which are not treated with NaOH, showed a higher concentration than 2000 mg/kg in the olive flesh. These results will contribute to the reevaluation of table olives from a nutritional and functional point of view because of the promising bioactivity properties attributed to olive triterpenic acids.     

Inhibitors of lactic acid fermentation in Spanish-style green olive brines of the Manzanilla variety

Frequently, a delay or lack of lactic acid fermentation occurs during the processing of Spanish-style green olives, in particular of the Manzanilla variety. Many variables can affect the progress of fermentation such as temperature, nutrients, salt concentration, antimicrobials in brines, and others. In this study, it was demonstrated that an inappropriate alkaline treatment (low NaOH strength and insufficient alkali penetration) allowed for the presence of several antimicrobial compounds in brines, which inhibited the growth of Lactobacillus pentosus. These substances were the dialdehydic form of decarboxymethyl elenolic acid either free or linked to hydroxytyrosol and an isomer of oleoside 11-methyl ester. Olive brines, from olives treated with a NaOH solution of low concentration up to 1/2 the distance to the pit, contained these antimicrobials, and no lactic acid fermentation took place in them. By contrast, a more intense alkaline treatment (2/3 lye depth penetration) gave rise to an abundant growth of lactic acid bacteria without any antimicrobial in brines. Therefore, the precise cause of stuck fermentation in Manzanilla olive brines was demonstrated for the first time and this finding will contribute to better understand the table olive fermentation process.     

A novel Lactobacillus pentosus-paired starter culture for Spanish-style green olive fermentation

A new starter culture consisting of two Lactobacillus pentosus strains was developed and successfully used for Spanish-style green olive fermentations in an industrial study. The inoculum, consisting of L. pentosus LP RJL2 and LP RJL3 strains, was inoculated in 10,000 kg glass fiber containers at 10⁶ CFU/ml and 10⁵ CFU/ml, final concentration respectively, in five different olive processing plants in the south of Spain. As a control, uninoculated fermentors were also used. In all inoculated fermentors, the paired starter rapidly colonized the brines to dominate the natural microbiota and persisted throughout fermentation. A decline in pH to reach about 5.0 was achieved in the first 15–20 days, reaching about 4.0 at the end of the process. The lactic acid concentration in brines increased rapidly in the first 20 days of fermentation (0.3–0.4 g/100 ml) to give values higher than 0.8 g/100 ml at the end of the process. In contrast, increasing lactic acid concentration was slower in uninoculated than in the inoculated brines, and the final concentrations were lower. Although reaching similar values at the end of the process, the decline in pH in uninoculated fermentors was slower than in the inoculated ones. These results show the efficacy of the new starter culture to control the lactic acid fermentation of Spanish-style green olives.     

Olive Fermentation and Processing: Scientific and Technological Challenges

ABSTRACT: The 3 main commercial table olive preparations are the Spanish-style green olives, the Greek-style naturally black olives, and the Californian-style black ripe olives. In all cases, fruits undergo fermentation in brine solution, which preserves them and increases palatability. Lactic acid bacteria dominate in brines of green olives, while mainly fermentative yeasts are found in brines of black ones. The fermentation is spontaneous and begins as soon as olives are put into brine. Research on table olives has been going on for 100 years, but many questions remain. Previous work has focused on the efficient utilization of starter cultures in order to control the fermentation, to guarantee the absence of spoilage, and to govern the relationship between olive flavor and microorganisms. Nowadays, there is concern over environmental problems that the fermentation solutions represent, in particular their high NaCl content. Fermentation or simply storing olives in water without salt presents some problems. The high concentration of sodium benzoate used for storing black ripe olives by the American industries is not advisable for European processes due to the residual amount of preservative in the final product. Fermentation of green olive wash waters, in order to obtain valuable products, is being pursued, but polymeric inhibitors are problematic and undesirable microorganisms grow. The consumption of organic foods is increasing and black olives fit the definition since they are processed without chemical treatment. However, the hydrolysis of the bitter compound oleuropein, either by microorganisms or acids, needs to be studied.     

A new polymer inhibitor of lactobacillus growth in table olives

The bulky precipitate formed in the washwaters from the processing of Spanish-style green olives, as a consequence of HCl acidification, presented inhibitory activity against lactic acid bacteria growth, the intensity of the effect being influenced by the type of washwater, pH and inoculum size. The delay or complete inhibition of microorganism growth was confirmed for three strains of lactobacilli (Lactobacillus pentosus LP99, L. pentosus ATCC 8041 and L. plantarum ATCC 14917), and this phenomenon can allow growth of undesirable microorganisms during washwater fermentation. The inhibitory activity against lactobacilli in acidified washwaters disappeared when the bulky precipitate was eliminated by centrifugation or simply by sonicating the solutions. It seems that sonication modified the macromolecular structure of the precipitate and the biosorption of microorganisms in it did not occur. Also, the inhibitory activity of this precipitate was confirmed in brines of Spanish-style green olives. Therefore, this new lactobacillus growth inhibitor will have to be taken into account in the future for improving olive fermentation.     

Content of biogenic amines in table olives

Content of biogenic amines in flesh and brines of table olives was determined by high-pressure liquid chromatography analysis of their benzoyl derivatives. No biogenic amines were found in the flesh of fresh fruits at any stage of ripeness. Contents of biogenic amines in Spanish-style green or stored olives increased throughout the brining period but were always higher in the former. Putrescine was the amine found in the highest concentration. Small quantities of cadaverine were found in the samples taken after 3 months of brining. This compound and histamine, tyramine, and tryptamine were also found in samples taken after 12 months. Gordal cultivar showed the highest contents, followed by Manzanilla and Hojiblanca. No relationship was found between contents of biogenic amines and lactic acid production or table olive spoilages, although zapatera olives had considerably higher amounts than those brines that had undergone a normal process. Concentrations in directly brined olives were markedly lower than contents in Spanish-style olives. With respect to partition between flesh and brine, there was equilibrium between both media in the case of Spanish-style olives, whereas the contents in directly brined olives were higher in flesh than brine.     

Occurrence of Listeria monocytogenes in green table olives

Microbiological safety of green table olives from different cultivars, prepared by both the Spanish-style and biological methods and fermented with starter cultures of lactic acid bacteria (Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus pentosus), was investigated. The fermentation process was monitored by measuring pH values, titratable acidities, and growth of lactic acid bacteria over time. During fermentation, lactic acid bacteria and yeasts were major microbial populations. Microbiological safety was evaluated by analysis for Listeria monocytogenes with the use of an enrichment method during storage (from 55 days to 18 months). Results demonstrated that L. monocytogenes can survive and grow in green table olives. L. monocytogenes was found in one of the commercial (thermally treated) samples analyzed and in all samples older than 2 months, irrespective of olive cultivar, lactic acid bacteria starter used, pH and titratable acidity of brine samples, or treatment applied.     

Effect of post-fermentation storage on Spanish-style green Manzanilla olives

The work studies the physicochemical, microbiological, and sensory changes which take place during the post-fermentation storage phase of Spanish-style green Manzanilla olives, achieved mainly by increasing NaCl to up to ∼90 g/L. The storage caused: i) an overall significant titratable acidity decrease and combined acidity increase, ii) a degradation of the texture of the fruits, iii) a considerable decrease in the lactic acid bacteria population, and iv) non significant changes in the yeast population. The later packaging of olives improved the colour index of the fruits, but did not prevent texture degradation. The multivariate statistical analysis of their sensory scores showed that the fermentation profiles of olives may persist even after the post-fermentation storage.     

Influence of processing, storage time, and pasteurisation upon the tocopherol and amino acid contents of treated green table olives

The effect of the processing system, storage time, and pasteurisation on the contents of tocopherols and individual amino acids was determined in alkali-treated green table olives. Olives were processed following three different systems (Spanish-style, short-process, and Picholine-style), together with two modified preparations of Spanish-style (without-washing process and olives packed with fermentation brine). Processing and pasteurisation had no effect on the -tocopherol and -tocopherol contents, but significant losses of both tocopherols were found after 12 months of storage. Calculation of protein quality of each product was made from its amino acid composition. The chemical score of treated green table olives, evaluated using the FAO reference protein for adults, ranged from 46 to 60%, the limiting amino acid being lysine. With the exception of lysine, amino acids can be considered to be stable during green olive processing, storage, and pasteurisation.     

Transformation of oleuropein and its hydrolysis products during Spanish-style green olive processing

Products obtained from elenolic acid glucoside hydrolysis during the storage of Spanish-style green olive brines were elenolic acid and glucose. The disappearance of elenolic acid glucoside from brines followed a first-order kinetics rate and was influenced by the storage temperature. Hydrolysis of this glucoside also occurred in brines ultrafiltered at 10000 Da pore size, indicating that no enzymatic action was needed to break the glycoside bond. Inoculation experiments with microorganisms obtained from a Spanish-style green olive brine demonstrated that this microflora was able to use the glucose formed from elenolic acid glucoside hydrolysis, although in our experiments only yeasts could grow in the assayed brines. It needs to be stressed that the glucose liberated by elenolic acid glucoside hydrolysis may represent about 20% of the total initial glucose in brines at the start of the fermentation step in Spanish-style green olive processing. © 1998 SCI.     

Survival of Bacillus cereus vegetative cells during Spanish-style fermentation of conservolea green olives

The aim of this research was to investigate the survival of artificially inoculated Bacillus cereus during Spanish-style green olive fermentation. Olives were initially treated with lye and subjected to different fermentation procedures including (i) heat shock (85 degrees C for 10 min) and inoculation with Lactobacillus plantarum ACA-DC 287, (ii) heat shock and inoculation with L. plantarum ACA-DC 146, (iii) heat shock and inoculation with Lactobacillus pentosus isolated during previous studies, and (iv) fermentation by indigenous flora (control process). Microbial growth and survival, pH, titratable acidity, and organic acid evolution were monitored. Inactivation of B. cereus was observed during all processes. The pathogen population declined during all fermentations, but a tailing effect was observed in the brines when the population reached 2 log CFU/ml, at which point the pathogen does not pose a risk to human health. The rate of inactivation was higher in heat-shocked inoculated olives (mean of -2.21 log CFU/day) compared with control olives (-1.26 log/day), indicating an advantage of heat shock and inoculation over spontaneous fermentation. The production of organic acids (primarily lactic acid) during fermentation seemed to be the main factor that determined the behavior of the pathogen under stress conditions prevailing in the brine. Principal components analysis was useful for distinguishing among the different fermentation processes on the basis of the relevant organic acid profile.     

Effects of alkali neutralization with CO2 on fermentation, chemical parameters and sensory characteristics in Spanish-style green olives (Olea europaea L.)

This paper aimed to study CO₂ use as alkali neutralizing agent instead of the traditional washings with water, in Spanish-style green olives. Titratable acidity, pH, sugars and microbial populations were monitored during processing. CO₂ treatment was effective in reducing water-soluble carbohydrate loss from the olives during washing step and affected combined acidity and lactic acid fermentation. The final pH of the fermented brine was higher than in control, mainly due to high buffering capacity of the solution. CO₂ treatment allowed the reduction of more than 80% of the wastewaters. Sensory evaluations of the end product were carried out. Olives treated with CO₂ were perceived to be more intense in acidic taste than control, while for texture attributes no differences were detected.     

Induced lactic acid fermentation of untreated green olives of the Conservolea cultivar by Lactobacillus pentosus

Inoculation of untreated green olives of the Conservolea cultivar (Olea europea media rotunda) with a commercial strain of Lactobacillus pentosus with/without glucose supplement was studied. Despite an initial loss in viability of 0.5 log cycles on average, due to lack of adaptation of the starter to the saline environment of the brine, cultures grew well and initiated an accelerated fermentation process. Inoculation reduced the survival period of Enterobacteriaceae, and consequently potential spoilage, and caused a quicker acidification of brines and decrease in pH compared with control uninoculated processes. The diffusion of phenolic compounds was slow, hindered by the epidermis of the fruits, and had no effect on the growth of the starter. HPLC analysis revealed that lactic and acetic acids were among the end‐products during the fermentation of green olives. Citric, tartaric and malic acids were also detected but at much lower concentrations. Results obtained from this work can be applied directly to industry for the effective use of starter cultures on natural fermentation processes of green olives without prior lye treatment. © 2003 Society of Chemical Industry     

Lactic acid bacteria from fermented table olives

Table olives are one of the main fermented vegetables in the world. Olives can be processed as treated or natural. Both have to be fermented but treated green olives have to undergo an alkaline treatment before they are placed in brine to start their fermentation. It has been generally established that lactic acid bacteria (LAB) are responsible for the fermentation of treated olives. However, LAB and yeasts compete for the fermentation of natural olives. Yeasts play a minor role in some cases, contributing to the flavour and aroma of table olives and in LAB development.     

Chloride salt mixtures affect Gordal cv. green Spanish-style table olive fermentation

This work studies the effects of different sodium (in the range of 4-10%), potassium (0-4%) and calcium (0-6%) chloride salt mixtures on the fermentation profile of Gordal olives processed according to the Spanish style. For this purpose, response surface methodology based on a simplex centroid mixture design with constrain (sum of salt percentages = 10%) was used. All treatments reached appropriate titratable acidity levels, but this parameter could not be related to the initial chloride salt concentration. The presence of CaCl₂ led to lower initial and after-fermentation pHs, delayed sugar diffusion into the brine, its maximum concentration and titratable acidity formation. CaCl₂ also delayed Enterobacteriaceae and yeast sprang, decreasing their overall growth. This chloride salt also showed a tendency to reduce overall lactic acid bacteria growth. KCl had a similar behaviour to NaCl but, in general, increased overall microbial growth. Thus, a partial substitution of NaCl in Spanish-style green olives with KCl and CaCl₂ does not substantially modify the fermentation profile but does produce some changes, which, when properly managed, could help to improve product processing.     

Influence of olive paste preparation conditions on virgin olive oil triterpenic compounds at laboratory-scale

The influence of olive paste preparation conditions on the triterpenic content of virgin olive oils from Arbequina and Picual cultivars was investigated. For this purpose, three sieve diameters of the hammer mill (4, 5, and 6 mm), two malaxation temperatures (20 and 30 °C), and two malaxation times (20 and 40 min) were tested. Results obtained showed that for Arbequina oils, a finer crushing level resulted in higher maslinic acid and erythrodiol content. Increasing malaxing temperature and time lead to a rise in both oleanolic and maslinic acid concentration, whereas erythrodiol content increased only for the longer malaxation time. For Picual oils, higher concentrations of oleanolic acid, maslinic acid, and uvaol were obtained by prolonging the paste malaxation time. A finer crushing level resulted also in an increase of maslinic acid content. These findings suggest that virgin olive oil triterpenic composition can be improved by regulating olive paste preparation conditions.     

Olive glutaraldehyde-like compounds against plant pathogenic bacteria and fungi

Aqueous solutions from table olives constitute a major environmental problem because of their high mineral and organic contamination but they also contain compounds with antimicrobial properties. In this study, the bactericidal and fungicidal activities of salt-free table olive solutions against several phytopathogenic micro-organisms were examined. The storage solutions of black ripe olives showed noticeable bactericidal activity against species of Erwinia, Clavibacter, Agrobacterium and Pseudomonas, although the washing waters from Spanish-style green olives were less effective. In fact, black olive storage solutions diluted to 20–30% were able to reduce the inoculated population of Erwinia amylovora and Pseudomonas syringae up to 6–7 log. This bactericidal activity was correlated with the presence of olive glutaraldehyde-like compounds such as the dialdehydic form of decarboxymethyl elenolic acid in these solutions, either free or linked to hydroxytyrosol. Experiments in vitro demonstrated that the bactericidal activity of these substances could explain most of the activity of the black olive storage solutions. Moreover, these salt-free table olive solutions also showed fungicidal activity against species of Phytophthora, Colletotrichum, Alternaria, Botrytis and Pestalotiopsis. The black olive storage solutions were more fungicidal than the washing solutions from Spanish-style green olives, with mycelial growth inhibition of up to 100% being attained, in some instances. These findings open the possibility of using table olive wastewaters in agriculture for pest management.     

Physicochemical, microbiological, and organoleptic profiles of Greek table olives from retail outlets

The physicochemical, microbiological, and organoleptic profile of different commercial table olive products from retail outlets was studied. Average pH values were 4.00, 3.96, and 4.31 for Spanish-style green, naturally black, and dry-salted olives, respectively, while salt content was 6.21, 7.34, and 8.00% for the same commercial products. Mean values for titratable acidity were 0.53 and 0.63% (wt/vol) for green and naturally black olives. In general, mean values for pH, titratable acidity, and salt content were in accordance with the requirements established by the International Olive Oil Council (IOOC) for the trade of table olives, although considerable variation was observed within individual olive samples. Salt content of dry-salted olives did not meet the minimum limit of 10% established by the IOOC. The dominant microbiota consisted of lactic acid bacteria and yeasts. Their population was less than 10(9) CFU ml(-1), as stipulated by the IOOC standard for fermented olives held in bulk in a covering liquid. These microorganisms come from the natural microbiota found in spontaneous fermentations and impose no risk to human health. No enterobacteria, pseudomonads, Bacillus cereus, or Clostridium perfringens were detected in any of the samples given the physicochemical characteristics found. The organoleptic profile varied greatly according to processing style and commercial preparation. Green olives had more uniform sensory characteristics than naturally black and dry-salted olives. The most important attributes that influenced the judgment of the panelists were salt content and crispness of the olives.     

天然食物中的抗艾滋病有效成分

天然食物中有抑制和阻断HIV感染的有效成分,如三萜类的齐墩果酸（Oleanolic acid）、熊果酸（Ursolic acid）、山楂酸（mastinic acid）等,黄酮类的金合欢素（acacetin）、芹菜素（apigenin）等,多糖类的芦荟多糖、海藻多糖、螺旋藻多糖等,蛋白类的苦瓜蛋白、乳铁蛋白等,发掘这些天然食物有效成分对艾滋病的防治具有重要意义.     

Use of selected autochthonous lactic acid bacteria for Spanish-style table olive fermentation

The present work presents a successful attempt to achieve an enhanced and more predictable fermentation process in Spanish-style green olive technology by selection and use of autochthonous starter cultures. During the first phase of this work, two Spanish-like fermentations of green table olives of cultivar (cv) "Nocellara del Belice", coming from irrigated and not irrigated fields, were monitored, in order to highlight the best agricultural conditions for drupe production and to isolate lactic acid bacteria strains with relevant technological properties. Among 88 identified isolates, one Lactobacillus pentosus strain showed remarkable biochemical features and high acidification rate in synthetic brine. In the second phase, the selected strain was used as starter culture in three different trials to establish the best conditions for its use. Microbial counting, as well as starter tracking by M13 RAPD-PCR, reflected the optimal adaptation of the strain to the environment. Spontaneous fermentation needed a 14-day long lag phase to reach the same population as the inoculated trials. Moreover, sensory traits of table olives obtained with adjunct culture showed better characteristics compared to those processed in the other trials, in particular concerning the presence of off-odours.