Bacteriocins (BCNs) are ribosomally synthesized polypeptides or proteins with antimicrobial activity, produced by different groups of bacteria. Many lactic acid bacteria (LAB) produce BCNs with broad spectra of inhibition. The antimicrobial activity of BCNs against spoilage organisms (SPOs) has raised considerable interest in their application in juice preservation. The objectives of the study were to: (i) isolate, identify and screen BCN producing bacteria for antimicrobial activity against spoilage bacteria and fungi, (ii) optimize production of BCN from selected producers and (iii) investigate the industrial application of the BCN as a preservative in fruit juice. Eleven LAB strains of BCN producers were screened for antimicrobial activity. BCNs from Lactobacillus plantarum and Pediococcus pentosaceus 34 were the most effective against juice spoilage bacteria and fungi. The effect of medium components on bacteriocin production in L. plantarum and P. pentosaceus 34 was also determined. Clementine:Valencia (1:1) juice was used for the first time as the growth medium for L. plantarum and P. pentosaceus 34. The BCN from L. plantarum showed the highest activity and was, therefore, chosen for juice fermentation studies. The identification of L. plantarum was confirmed by biochemical tests, polymerase chain reaction (PCR) and sequencing of the recA gene. The highest BCN activity was observed for L. plantarum grown in De Man-Rogosa-Sharpe (MRS) and a combination of all supplements (i.e. peptone, MnSO4.H2O, Tween 80, glucose and whey), followed by MRS and Tween 80, peptone, MnSO4.H2O and MRS alone. MRS was a better medium for BCN production than juice [Clementine:Valencia (1:1)]. Size exclusion chromatography (SEC) was used to isolate the active L. plantarum BCN fraction which corresponded to an approximate molecular weight of 3.2 kDa and was proteinaceous in nature. Plantaricin structural genes (plnEF, plnJ, plnK, plnN) were detected in the L. plantarum strain by PCR and sequenced, and were chromosomally encoded as no plasmids could be detected. This implies that the BCN from L. plantarum is most likely a type of class IIa plantaricin which is responsible for the broad inhibitory activity observed. For the industrial application studies, L. plantarum BCN-containing cell free supernatant (BCNsup) added to “Ready to Drink” (RTD) Clementine:Valencia (1:1) juice at concentrations of 3 600 - 500 000 ppm decreased growth of SPOs, Lactobacillus acidophilus and Streptococcus thermophilus. At 250 000 ppm, the L. plantarum BCNsup achieved 5.3 and 6.8 log reductions of the L. acidophilus, after 24 and 48 h, respectively, which is larger than the USFDA (2001) requirement of a 5 log reduction in SPO activity, for preservation of fruit juices. However, there was a decrease in the activity when the BCNsup was applied to industrial (Valor) RTD juice (mango-orange) at decreasing concentrations of 100 000, 50 000 and 25 000 ppm. Organoleptic tests showed that the BCN did not alter flavor or taste of the juice and did not cause toxicity or allergic reactions. A food safety risk assessment was conducted in order to determine the Critical Control Point(s) [CCP(s)] at which the BCN could be applied to control identified microbiological hazards, and a Hazard Analysis and Critical Control Point (HACCP) plan was developed. This is the first report on the optimisation of L. plantarum BCN production in juice [Clementine:Valencia (1:1)], followed by inoculation into RTD juice (mango-orange), including a HACCP plan for the application of the BCN as a preservative in juice.

Thermal processing is the primary mode of preservation of mango products, which can be considerably damaging to the delicate quality of mango. HP processing could thus be a potential alternative for extending the shelf-life of mango products. Establishing HP processing technology need data on microbial inactivation kinetics and shelf-life study of the product. The objective of this research was therefore to evaluate the application of HP treatment for inactivation of microorganisms (pathogenic and spoilage type) and to evaluate the shelf-life of HP treated mango juice. / HP destruction kinetics of three common spoilage microorganisms, Leuconostoc mesenteroides, Zygosaccharomyces bailii and Pichia membranaefaciens and two pathogenic microorganisms, Escherichia coli O157:H7 and Listeria monocytogenes Scott A, were evaluated at 250-550 MPa with 0-60 min holding time at room temperature with species specific initial counts between 10 6 to 108 CFU/mL.

No description available.

Orientador: Fumio Yokoya / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-22T20:33:42Z (GMT). No. of bitstreams: 1 Conti_MariaJosiane_M.pdf: 10300128 bytes, checksum: c571b2db431105550b581c8203ff0630 (MD5) Previous issue date: 2013 / Resumo: O Alicyclobacillus acidoterrestris, é uma bactéria acidotermofílica, não patogênica, de origem ambiental. Este organismo é um potencial agente deteriorante em polpas e sucos de frutas ácidos. Devido à presença de endosporos de elevada resistência térmica, torna-se difícil sua inativação pelos tratamentos térmicos convencionais de pasteutização. Esta pesquisa visou extrair e processar termicamente o suco de seis tipos de frutas tropicais, de caráter ácido (abacaxi, acerola, caju, goiaba, manga e maracujá), e avaliar se estes apresentam condições para que o A. acidoterrestris se desenvolva em sua temperatura ótima (45°C) e também em temperatura de abuso de processamento (35°C), temperatura que favorece o crescimento de microrganismos mesófilos. Além disso, foi analisada a concentração mínima inicial, do microrganismo em estudo, capaz de sobreviver e se desenvolver no suco de manga, aquele que apresentou melhores condições de desenvolvimento na primeira fase do estudo. Neste estudo foi também incluída a análise qualitativa de guaiacol como indicador de deterioração. Foi preparada uma suspensão com a mistura de esporos de quatro cepas de A. acidoterrestris em mesma concentração, padronizada e inoculada nos sucos, previamente extraídos e tratados termicamente, apresentando carga inicial no suco de 102 UFC/mL. O desenvolvimento do microrganismo foi acompanhado através de plaqueamentos em meio de cultura próprio, por até 30 dias, ou até que atingissem uma contagem constante. Ao final, foram avaliados o desenvolvimento máximo da bactéria, em cada suco, na respectiva condição de cultivo, e o período que levou para atingir estes determinados valores. Apenas no suco de caju o pool de bactérias não apresentou desenvolvimento, no prazo máximo estipulado do ensaio (30 dias). O microrganismo apresentou desenvolvimento em todos os demais sucos, alcançando índices de produção de guaiacol e outros subprodutos / Abstract: Alicyclobacillus acidoterrestris is an acidophilic bacterium, nonpathogenic, of environmental origin. This organism is a potential spoiler acid frutits, pulps and juices. Due to the presence of endospores of high heat resistance, it becomes difficult to inactivate it by pasteurization conventional thermal treatment. This research aimed to extract and process thermally the juice of six kinds of tropical fruit, acid character (pineapple, acerola, cashew, guava, mango and passion fruit) and to evaluate if these have conditions for Alicyclobacillus acidoterrestris to develop in optimum temperature (45°C) and also in processing abuse temperature (35°C). Besides, it was analyzed the minimum initial concentration of the microorganism under study, which is able to survive and develop in the mango juice, which presented better conditions of development, according to the first phase of the study. A suspension was prepared with the spores mixture of four strains of A. acidoterrestris in the same concentration, because in preliminary studies it was possible to verify that some strains did not develop in all types of juice. The suspension was standardized and inoculated into the previously extracted and heart treated juices, showing initial concentration of 102 CFU/mL in the juice. The development of the microorganism was accompanied through plating in specific culture medium, for around 30 days or until it reached a constant counting. In the end, bacterium maximum growth was evaluated in each juice, in its respective growing condition and the time it took to achieve these values. The pool of bacteria didn't grow only in the cashew juice. All other juices showed development of the microorganism, reaching production rates of guaiacol and other byproducts, in accordaner with to previous studies / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos