Lactic acid bacteria produce pediocin-like bacteriocins designated as Class Ha. These antimicrobial peptides are antagonistic against Listeria monocytogenes and other closely related Gram-positive bacteria Self-protection of the producer organism is attributed to the immunity proteins, encoded by genes that are eo-transcribed with the structural gene that encode the bacteriocin. The lactic acid bacterium, Leuconostoc gelidum UAL 187-22 is immune to its own bacteriocin, leucocin A. This is accredited to its immunity protein and the possible absence of a receptor on its cytoplasmic membrane. Leucocin A was purified from the supernatant of 1. gelidum to 90% purity by ion-exhange chromatography and C18 reverse phase High Pressure Liquid Chromatography (RP-HPLC) eluted with an acetonitrile, 0.1% Triflouroacetic acid (TFA) gradient. The immunity gene was isolated from the same producer using the polymerase chain reaction from the recombinant plasmid pJF 5.5 using primers EAL-2 and EAL-3. The amplicon was truncated into versions A and B by removing the C- and N-terminals, with HaeIII and ClaI restriction enzymes, respectively. The amplicon and the truncated fragments A and B were cloned into pMALc2 to construct recombinant plasmids pKPl, pKPIA and pKPIB, correspondingly, which were transformed into Escherichia coli (E. coli) strain JMI03. Clones were confirmed by colony PCR and Southern blot hybridization. The recombinant clones were subsequently expressed as MBP-IP, MBP-IPA and MBP-IPB fusion proteins that were verified by Western blot using the anti-MBP antibody. Factor Xa protease was used to cleave MBP from the proteins of interest. The resulting pure immunity protein versions had an approximate molecular weight of slightly more that 10 kDa. The binding interactions of the purified immunity protein constructs and leucocin A were compared on the Biacore 2000 instrument with surface plasmon resonance. None of the immunity constructs interacted with leucocin A, however, the N-terminal region of the immunity protein interacted with the cytoplasmic extract. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

The blending of lignin as a component in a thermoplastic blend poses a challenge in the form of dispersion and compatibility. Polyesters such as poly(lactic acid) and poly(butylene adipate-co-terephthalate) offer the best opportunity of compatibility in melt blending with lignin due to their ability to form hydrogen bonds. The fractionation of lignin into more homogeneous fractions offers better dispersion and more consistent properties, retaining the toughness of the original polymer in addition to bridging stress transfer between PLA and PBAT. Functionalization of lignin was done by lactic acid grafting. The resulting blend of PLA/PBAT/modified fractionated lignin showed improved interaction between lignin and PLA, but reduced compatibility between lignin and PBAT. This thesis provides a deeper understanding on the effect of lignin heterogeneity, its fractions, and the functionalization of lignin on lignin and bioplastic blends to further the use of a largely produced industrial by-product in high value applications. / Natural Sciences and Engineering Research Council (NSERC) – Lignoworks Biomaterials and Chemicals Strategic Research Network, Canadian Foundation for Innovation (CFI), Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA)

To gain a better understanding of contaminating bacteria in bioethanol industry, we profiled the bacterial community structure in corn-based bioethanol fermentations and evaluated its correlation to environmental variables. Twenty-three batches of corn-mash sample were collected from six bioethanol facilities. The V4 region of the collective bacterial 16S rRNA genes was analyzed by Illumina Miseq sequencing to investigate the bacterial community structure. Non-metric multidimensional scaling (NMDS) ordination plots were constructed to visualize bacterial community structure groupings among different samples, as well as the effects of multiple environmental variables on community structure variation. Our results suggest that bacterial community structure is facility-specific, although there are two core bacterial phyla, Firmicutes and Proteobacteria. Feedstock, facility, and fermentation technology may explain the difference in community structure between different facilities. Lactic acid, the most important environmental variable that influences bacterial community structure grouping, could be utilized as an indicator of bacterial contamination. We also identified genes responsible for the multiple antibiotic-resistance phenotype of an Enterobacter cloacae strain isolated from a bioethanol fermentation facility. We performed PCR assays and revealed the presence of canonical genes encoding resistance to penicillin and erythromycin. However, a gene encoding resistance to virginiamycin was not detected.

The purpose of this study was to examine VT and LT in boys and men. Eight boys (10-11 years) and nine men (18-30 years) completed a graded exercise test on a cycle ergometer. A two-way (group x threshold) ANOVA compared physiological responses (V02 1/min and ml/kg/min, percentage of V02max, and HR) at VT and LT. Statistical significance was set at P < 0.05. No significant interaction was observed. Significant main effects for group included a higher V02 (1/min) in the men, and a higher percentage of VO2max in the boys. Significant main effects for threshold showed all variables were greater at VT than LT. Within each group, all variables were significantly higher at VT than LT. For the boys and all subjects together, significant correlations between thresholds were observed for V02 (1/min and ml/kg/min) and HR, but not percentage of VO2max. For the men alone, no significant correlations were found. Together, these results suggest that physiological changes associated with LT may contribute to the onset of VT, and the occurrence of the thresholds (expressed as a percentage of VO2max) declines with maturation. / School of Physical Education

L(+)-Lactic acid is a commonly occurring organic acid, which is valuable due to its wide use in food and food-related industries, and its potential for the production of biodegradable and biocompatible polylactate polymers. The aim of this study was to optimize and scale-up a biotechnological process of L(+)-lactic acid production by suspended cells of R. arrhizus DAR 36017 with waste potato starch as the substrate. Commonly used inorganic and organic nitrogen sources, including ammonium sulphate, ammonium nitrate, urea, yeast extract and peptone, were assessed in conjunction with various ratios of carbon to nitrogen (C:N). Fermentation media with a low C:N ratio enhanced the production of lactic acid, biomass and ethanol, while a high C:N ratio led to production of more fumaric acid as a by-product. The use of organic nitrogen sources (yeast extract, peptone and urea) resulted in a significant reduction of lactic acid yields by 15% - 34% with a decrease of C:N from 168 to 28. The use of inorganic nitrogen sources (ammonium nitrate and ammonium sulphate) led to a high lactic acid yield of 84% - 91% at a C:N below 168. Therefore, ammonium nitrate and ammonium sulphate were considered to be better nitrogen sources for lactic acid production. Small pellets are the favoured morphological form for many fermentation processes by filamentous fungi. However, to control filamentous Rhizopus sp in the pellet form in a submerged fermentation system is difficult due to its filamentous characteristics. An acidadapted preculture technique was developed to induce the formation of the pellet form in bioreactors. Using the acid-adapted precultures, the fungal biomass can be controlled in small dispersed pellets as a dominant morphological form. With these small pellets, a lactic acid yield of 86-89%, corresponding to a concentration of 86-89g/L, was obtained in a laboratory scale process using a stirred tank reactor (STR) and a bubble column reactor (BCR). A batch bioprocess for lactic acid production was successfully scaled-up from shake flasks to laboratory scale bioreactors. Results from a simulated scale-up process revealed that the concentration and productivity of lactic acid decreased with the increase of the scale-up steps because of increased pellet size. This suggested that a one-step scale-up process using the acid-adapted preculture may be feasible in an industrial-scale bioreactor system. A comprehensive investigation of the impact of cultivation parameters on the morphology of R. arrhizus and lactic acid production was carried out in the BCR. The results showed that the fungal morphology was significantly influenced by carbon sources, pH, starch concentrations, sparger designs and aeration rates. The favoured morphology for lactic acid production was freely dispersed small pellets, which could be retained as a dominant morphology under operation conditions at pH 5.0 – 6.0, starch concentrations of 60 – 120 g/L and aeration rates of 0.2 – 0.8 vvm, using a sintered stainless steel disc sparger. The optimal cultivation conditions at pH 6.0 and aeration rate of 0.4 vvm resulted in the formation of the freely dispersed small pellets and production of 103.8 g/L lactic acid, with a yield of 87%, from 120 g/L liquefied potato starch in 48 h. This study shows a technically feasible and economically promising process for the production of lactic acid from waste potato starch. The use of waste potato starch instead of pure glucose or starch as substrate can significantly reduce the production cost, making this technology environmentally and economically attractive. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1339122 / Thesis (Ph.D.) -- University of Adelaide, School of Chemical Engineering, 2008

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2005. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser.

Thesis (Ph.D.)--Mississippi State University. Department of Food Science, Nutrition and Health Promotion. / Title from title screen. Includes bibliographical references.

Thesis (MSc Food Sc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: A wide variety of traditionally and commercially fermented milks are commonly consumed in various countries of Sub-Saharan Africa. Commercially fermented milk is produced on an industrial scale according to well-managed, standardised production processes and starters are used to initiate fermentation. Traditionally fermented milk is prepared domestically and fermentation occurs spontaneously at ambient temperatures. Lactic acid bacteria (LAB) are responsible for milk fermentation during which they convert the milk carbohydrates to lactic acid, carbon dioxide, alcohol and other organic metabolites. Acetic acid bacteria (AAB), yeasts and mycelial fungi have also been isolated from fermented milks. In this study the microbial consortium present in three traditionally fermented milks, namely omashikwa from Namibia, masse from Mozambique and chekapmkaika from Uganda and two commercially fermented milks, namely chambiko from Malawi and omaere from Namibia, were isolated and enumerated on six different selective media that included MSR + C (specific for lactobacilli), KCA + TTC (specific for lactococci), KCA + V (specific for leuconostocs), MRS + E (specific for AAB), MEA (specific for mycelial fungi) and YPD (specific for yeasts). No significant differences were found between the enumeration values obtained for the three chambiko samples, as well as for enumeration values obtained for the two omaere samples on each of the selective media, indicating low sample variance. Significant differences between enumeration values obtained for the three omashikwa samples were found on all six selective media. Significant differences between enumeration values of the three masse samples and both the chekapmkaika samples were also observed on the selective media. In addition to this, significant differences were observed between average enumeration values obtained for each media between the masse and chekapmkaika, the chambiko and omaere, as well as when the traditional and commercial milks were compared. According to the average enumeration values obtained on each media selective for LAB, the highest bacterial counts were detected on KCA + TTC medium for omaere (2.3 x 106 cfu.ml-1), KCA + V for chambiko (1.8 x 105 cfu.ml-1), KCA + TTC for omashikwa and MRS + C for masse and chekapmkaika (6.2 x 106 and 2.0 x 103 cfu.ml-1, respectively). After isolation and enumeration of the microbes present in each milk, bacterial isolates on the media selective for LAB and AAB were obtained according to the Harrison Disk method. These isolates were identified by amplifying a 1.5 kilobase (kb) part of the 16S ribosomal RNA (rRNA) gene using the polymerase chain reaction (PCR), followed by DNA sequencing. The isolates were identified by comparing the sequences obtained to sequences listed in the NCBI database using the BLAST algorithm and searching for the closest relative. The main LAB group present in the omaere was lactococci (94%), in chambiko and chekapmkaika it was lactobacilli (30% and 45%, respectively), in omashikwa it was enterococci (43%) and in masse it was leuconostocs (68%). The same microbial species were present on a number of the selective media used in this study. Lactococcus spp., Enterococcus spp. and Lactobacillus spp. were isolated from MRS + C, KCA + TTC, KCA + V and MRS + E and Leuconostoc spp. were isolated from MRS + C, MRS + E and KCA + V. Hygienic standards during traditional milk fermentation is often poor and, therefore, microbial contaminants were isolated from the traditional milk and these included Acinetobacter johnsonii and Klebsiella pneumoniae from KCA + V, Mesorhizobium loti, Acinetobacter radioresistens, Escherichia coli, Staphylococcus spp., Kluyvera georgiana, Enterobacter spp. and Klebsiella oxytoca from KCA + TTC, Staphylococcus spp. from MRS + C and Bacillus spp. from MRS + E. Since the media used for the isolation of the LAB and AAB in this study were not selective further identification of the enumerated microbes is of importance for the identification of the microbial groups present in each fermented milk. The data obtained in this study clearly shows that fermented milks from Sub-Saharan Africa vary significantly from each other in terms of microbial numbers, microbial diversity and the dominant microbial groups present. The microbial diversity of the traditionally fermented milks was more diverse than the microbial diversity of the commercially fermented milks. LAB strains isolated from these traditionally fermented milks can be used to develop novel starters and as a result new commercially fermented dairy products with unique aromas, tastes and characteristics can be produced. / AFRIKAANSE OPSOMMING: 'n Wye verskeidenheid tradisioneel en kommersieel gefermenteerde melk produkte word algeneem verbruik in verskeie lande van Sub-Sahara Afrika. Kommersieel gefermenteerde melk word geproduseer op groot skaal, deur deeglik bestuurde gestandardiseerde produksieprosesse en 'n beginkultuur word gebruik om fermentasie te inisieer. Tradisioneel gefermenteerde melk word tuis gemaak en fermentasie gebeur spontaan by kamertemperatuur. Melksuurbakterieë (MSB) is verantwoordelik vir melkfermentasie waartydens die bakterieë koolhidrate omskakel na melksuur, koolstofdioksied, alkohol en ander organiese sure. Asetaatsuurbakterieë (ASB), giste en miseliale fungi is ook al van gefermenteerde melk geïsoleer. In hierdie studie is die mikrobiese konsortium teenwoordig in drie soorte tradisioneel gefermenteerde melk, naamlik omashikwa van Namibië, masse van Mosambiek en chekapmkaika van Uganda en twee soorte kommersieel gefermenteerde melk, naamlik chambiko van Malawi en omaere van Namibië, geïsoleer en getel op ses verskillende selektiewe groeimedia insluitend MRS + C (spesifiek vir lactobacilli), KCA + TTC (spesifiek vir lactococci), KCA + V (spesifiek vir leuconostocs), MRS + E (spesifiek vir ASB), MEA (spesifiek vir miseliale fungi) en YPD (spesifiek vir giste). Geen betekenisvolle verskille is gevind tussen die mikrobiese tellings verkry vir die drie chambiko monsters nie, sowel as tussen die mikrobiese tellings verkry vir die twee omaere monsters, op elk van die selektiewe groeimedia, wat dui op lae monster variansie. Betekenisvolle verskille is gevind tussen die mikrobiese tellings verkry vir die drie omashikwa monsters op al ses selektiewe groeimedia. Betekenisvolle verskille is ook waargeneem tussen die mikrobiese tellings van die drie masse monsters en beide die chekapmkaika monsters op die selektiewe groeimedia. Daarbenewens is betekenisvolle verskille waargeneem tussen gemiddelde mikrobiese tellings verkry vir elke groeimedium tussen die masse en chekapmkaika, die chambiko en omaere asook toe die tradisionele en kommersiële melk produkte met mekaar vergelyk is. Volgens die gemiddelde mikrobiese tellings verkry op elk van die groeimedia selektief vir MSB, is die hoogste mikrobiese telling waargeneem op KCA + TTC medium vir omaere (2.3 x 106 kve.ml-1), KCA + V vir chambiko (1.8 x 105 kve.ml-1), KCA + TTC vir omashikwa en MRS + C vir masse en chekapmkaika (6.2 x 106 en 2.0 x 103 kve.ml-1, respektiewelik). Na die isolasie en tel van die mikrobes teenwoordig in elke melk is bakteriese isolate op die media selektief vir MSB en ASB verkry volgends die Harrison Disk metode. Hierdie isolate is geïdentifiseer deur amplifikasie van „n 1.5 kilobasis (kb) gedeelte van die 16S ribosomale RNS (rRNS) geen deur gebruik te maak van die polimerase kettingreaksie gevolg deur DNS klonering. Die isolate is geïdentifiseer deur die gekloneerde insetsels se volgordes te vergelyk met volgordes beskikbaar op die NCBI webwerf deur van die BLAST algoritme gebruik te maak en die naas verwante insetsel op te spoor. Die hoof MSB groep teenwoordig in die omaere was lactococci (94%), in chambiko en chekapmkaika was dit lactobacilli (30% en 45%, respektiewelik), in die omashikwa was dit enterococci (43%) en in die masse was dit leuconostocs (68%). Dieselfde mikrobiese spesies was teenwoordig op verskeie van die selektiewe groeimedia gebruik in hierdie studie. Lactococcus spp., Enterococcus spp. en Lactobacillus spp. is geïsoleer van MRS + C, KCA + TTC, KCA + V en MRS + E en Leuconostoc spp. is geïsoleer van MRS + C, MRS + E en KCA + V. Higiëniese standaarde tydens tradisionele melkfermentasie is dikwels swak en dus is mikrobiese kontaminante geïsoleer van die tradisionele melk produkte insluitend Acinetobacter johnsonii en Klebsiella pneumoniae van KCA + V, Mesorhizobium loti, Acinetobacter radioresistens, Escherichia coli, Staphylococcus spp., Kluyvera georgiana, Enterobacter spp. en Klebsiella oxytoca van KCA + TTC, Staphylococcus spp. van MRS + C en Bacillus spp. van MRS + E. Aangesien die media wat gebruik is vir die isolasie van die MSB en ASB in hierdie studie nie selektief was nie, is verdere identifikasie van die getelde mikrobes belangrik vir die identifikasie van die mikrobiese groepe teenwoordig in elke melk. Die data verkry in hierdie studie dui aan dat gefermenteerde melk produkte van Sub-Sahara Afrika betekenisvol van mekaar verskil in terme van mikrobiese getalle, mikrobiese diversiteit en die dominante mikrobiese groepe teenwoordig. Die mikrobiese diversiteit van die tradisioneel gefermenteerde melk produkte was meer divers as die mikrobiese diversiteit van die kommersieel gefermenteerde melk produkte. MSB spesies geïsoleer van hierdie tradisioneel gefermenteerde melk produkte kan gebruik word om nuwe beginkulture te ontwikkel en gevolglik kan nuwe kommersieel gefermenteerde suiwelprodukte met unieke aromas, smake en eienskappe geproduseer word.