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Thesis (MSc) -- University of Stellenbosch, 2005. / ENGLISH ABSTRACT: The Rooibos tea plant (Aspalathus linearis) is indigenous to South Africa and occurs only in the Western Cape's Cedarberg region. Rooibos tea is produced from the leaves and fine stems of the plant. The tea is normally prepared by brewing the leaves and consuming the liquor. However, the Rooibos plant is not only used to prepare tea; the plant extracts are also used in various neutraceutical and pharmaceutical products, including health drinks, iced tea, soaps and moisturising creams. Although the tea plant contains native enzymes responsible for the colour and aroma development of Rooibos tea, the disruption and maceration of the plant material during processing is insufficient to allow these enzymes proper access to the substrates responsible for Rooibos tea's characteristics. The current processing of Rooibos tea is also time consuming and is done under uncontrolled conditions, leading to unnecessary loss in aroma and antioxidant content. The addition of enzymes could improve the maceration of the plant material, shorten the processing time and improve the extraction of aroma, colour and antioxidant components. During this study, 16 commercially available microbial enzymes were evaluated on three different Rooibos substrates for the improvement of aroma and colour development, as well as the extraction of soluble solids (SS) and total polyphenols (TP). Thirteen enzymes were evaluated on spent tea for the enhanced extraction of soluble solids and to determine the best candidates for further evaluation on fermented and green Rooibos tea. Seven of the enzymes improved the yield in SS from spent tea. Up to 232% improvement was obtained, depending on the type of enzyme and dosage applied. The best six enzyme preparations were further evaluated on fermented Rooibos tea. For Depol™ 670L at 20 ul/g tea, the laboratory treatment increased the yield in SS by 44%, while small-scale industrial simulations increased the SS by 26%. However, an increase in the yield in SS was usually accompanied by a decrease in the %TP/SS ratio, indicating that mainly inactive compounds were extracted. Based on the results with the commercial enzymes, twelve "synthetic" enzyme cocktails, consisting of different combinations of commercial enzymes were designed, of which three cocktails released increased amounts of SS without decreasing the %TP/SS ratio significantly. Thirteen enzymes were evaluated on dried and freshly cut green Rooibos tea, with three enzymes (Depol™ 670L, Pectinex Ultra SP-L and Depol™ 692L) increasing the yield in SS between 21% and 66%, and the TP content between 11% and 47%. Laccase was the best candidate in improving colour development from green tea, with the improvement being slightly better at 50°C than at 40°C. All the "synthetic" cocktails containing laccase improved the colour extract of all three substrates evaluated, but also significantly decreased the TP and antioxidant content. However, lower dosages of laccase resulted in colour development with little loss in the antioxidant content. Due to the promising results obtained with the treatments of Rooibos tea with laccases, it was decided to clone and express the laccase gene (lacA) of Pleurotus ostreatus into Aspergillus niger. The gene was successfully transformed into A. niger, but the expression of the recombinant gene was not effective. / AFRIKAANSE OPSOMMING: Die Rooibostee plant (Aspalathus linearisi is inheems tot Suid-Afrika en kom slegs in die Sederberg-omgewing in die Wes-Kaap voor. Rooibostee word van die blare en fyn stingels van die plant geproduseer. Die tee word normaalweg voorberei deur die blare in kookwater te laat trek en dan die aftreksel te drink. Die Rooibos plant word nie net gebruik om tee te maak nie; die tee ekstrak word ook gebruik vir verskeie neutraseutiese en farmaseutiese produkte, insluitende gesondheidsdrankies, ystee, seep en bevogtigingsrome. Ten spyte daarvan dat die teeplant sy eie ensieme vir die kleur en aroma ontwikkeling van Rooibostee bevat, is die verbreking en maserasie van die plantmateriaal tydens prosessering onvoldoende om die ensieme genoeg toe gang tot die substrate verantwoordelik vir die kenmerkende eienskappe van Rooibostee te gee. Die huidige prosessering van Rooibostee is ook tydrowend en geskied onder onbeheerde toestande, wat tot 'n onnodige verlies in aroma en antioksidante lei. Die toevoeging van ensieme kan die afbraak van die plantmateriaal verbeter, die behandelingsproses verkort en die aroma, kleur en antioksidant inhoud van ekstrakte verbeter. Tydens hierdie studie is 16 kommersieel-beskikbare mikrobiese ensieme op drie verskillende Rooibos substrate vir die verbetering van aroma, kleur en ekstraksie van oplosbare vastestowwe (SS) en totale polifenole (TP) getoets. Dertien ensieme is op oorskot tee vir die verbeterde ekstraksie van oplosbare vastestowwe geevalueer, waama die beste kandidate vir evaluering op gefermenteerde en ongefermenteeede Rooibostee gekies is. Sewe ensieme het die SS vanaf oorskot tee verhoog. Tot 232% verhoging is waargeneem, afhangende van die tipe ensiem en die dosis wat gebruik is. Die beste ensiern preparate IS verder op gefermenteerde Rooibostee geevalueer, Labarotoriurn behandelings met Depol™ 670L teen 20 ul/g tee het die SS inhoud met 44% verhoog, terwyl die kleinskaalse industriele simulasie die SS inhoud met 26% verhoog het. 'n Verhoging in SS het egter gewoonlik met 'n afname in die %TP/SS verhouding gepaard gegaan, wat aandui dat hoofsaaklik onaktiewe stowwe vrygestel IS. Na aanleiding van die resultate met die kommersiele ensieme, is twaalf "sintetiese" ensiemmengsels met verskillende ensiemkombinasies getoets, waarvan drie mengsels ook meer SS vrygestel het met byna geen verlaging in die %TP/SS verhouding nie. Dertien ensieme was op gedroogde en vars gekerfde groen Rooibostee getoets met drie ensieme (Depol™ 670L, Pectinex Ultra SP-L en Depol™ 692L) wat die SS met tussen 21% en 66%, en die TP inhoud met tussen 11% en 47% verhoog het. Lakkase was die beste kandidaat vir die verbetering van kleur ontwikkeling by groen Rooibostee met die verbetering effens beter by 50°C as by 40°C. Al die "sintetiese" ensiem mengsels wat lakkase bevat het, het die kleur by al die verskillende substrate verbeter, maar het ook die TP en antioksidant inhoud aansienlik verlaag. Laer lakkase dosisse het goeie kleurontwikkeling tot gevolg gehad met minimale verlies in die antioksidant inhoud. Vanwee die goeie resultate wat met die lakkase behandelings verkry is, is daar besluit om die lakkase geen (lacA) van Pleurotus ostreatus te kloneer en in Aspergillus niger uit te druk. Die geen is suksesvol in A. niger getransformeer, maar die uitdrukking daarvan was nie effektief nie.

Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Aspalathus linearis is a leguminous shrub native to the Cedarberg Mountains in the Western Cape, of which the leaves and stems are used for the preparation of rooibos tea. Over the past few decades, rooibos tea and other related products have gained popularity due to their health promoting properties. These beneficial properties can partly be ascribed to the phenolic constituents that are trapped within the cellulolytic plant material of the tea leaves as glycoconjugated aroma and phenolic compounds. Although many fungal species are known for their efficient hydrolysis of plant material, fungal enzymes have not been evaluated for the bioprocessing of rooibos tea to improve its commercial value. It was the objective of this study to identify a specific cocktail of microbial enzymes to enhance the maceration of the rooibos plant material, while retaining the antioxidant content. During this study, 11 fungal species known for the production of hydrolytic enzymes, as well as 12 species isolated from rooibos tea products, were screened for their potential to improve aroma development and/or increased extraction of soluble matter and/or antioxidants from rooibos tea material. After culturing in Potato Dextrose medium, the crude enzyme extracts of the 23 isolates were evaluated on spent rooibos tea for enhanced extraction of soluble solids (SS) and/or total polyphenols (TP). Nine strains increased the yield in SS (improvement varying from 3% to 42%), while 14 strains yielded higher levels of TP (increase varying from 1% to 36%). Little improvement in colour development from green (unfermented) rooibos tea was observed, but the enzyme extracts from Pleurotus ostreatus var. florida, Lentinula edodes, Aspergillus oryzae, Aspergillus tubingensis, Paecilomyces variotti and Trichoderma reesei improved the aroma development from green tea to some extent. Ten-fold concentrated enzyme extracts from four of these isolates were able to release at least an additional 10% in SS from the green tea. The crude enzyme extracts prepared from three food-grade strains, i.e. Aspergillus oryzae, Lentinula edodes and Pleurotus ostreatus var.florida, contained relatively high levels of endoglucanase, xylanase and pectinase activities. Eight different culture media were evaluated for optimal hydrolase and laecase production by these food-grade fungi. MYPG proved to be the best growth medium, while 1% spent grain, 1% wheat straw and 1% pineapple peel gave the best induction of xylanase, cellulase, pectinase and laecase activities for L. edodes. When cultured in the Yeast Peptone (YP) medium + 1% wheat straw, the L. edodes enzyme cocktail showed the best improvement in both the aroma and colour development of green tea and may be considered for shortening of the fermentation time required for green tea processing. Traditional open-air fermentation of rooibos tea can take up to -1-6hours, which results in a significant loss in antioxidants and therefore also in its pharmaceutical and nutraceutical value. The Rhizopus oryzae cocktail prepared in YP + 1% wheat straw showed potential for the development of a quick-draw fermented tea made by infusion, where there is improved colour release and more than 20% improved extraction of soluble solids without a loss in the TP content. When cultured in Potato Dextrose medium, the L. edodes cocktail can be used for aroma and colour development from green tea, while the R. oryzae cocktail can be used for increasing the antioxidant content in rooibos extracts from green or fermented tea. This was confirmed with small-scale industrial treatments of fermented tea where the L. edodes YP + wheat straw cocktail improved the release in SS by more than 10% and the R. oryzae yP + wheat straw cocktail increased the yield in SS by more than 30% and the TP by more than 20%. / AFRIKAANSE OPSOMMING: Aspalathus linearis is 'n fynbosplant inheems aan die Sederberge in die Wes-Kaap, waarvan die blare en stingels vir die voorbereiding van rooibostee gebruik word. Die afgelope paar dekades het die gewildheid van rooibostee en verwante produkte aansienlik toegeneem weens die gesondheidsvoordele wat dit inhou. Hierdie voordelige eienskappe kan toegeskryf word aan die fenoliese komponente wat binne die sellulolitiese plantweefsel van die teeblare as gekonjugeerde geur- en fenoliese verbindings vasgevang is. Alhoewel verskeie swamspesies vir hul doeltreffende degradering van plantmateriaal bekend is, is fungale ensieme nog nie geëvalueer vir die prosessering van rooibostee om die kommersiële waarde daarvan te verbeter nie. Die doelwit van hierdie studie was om 'n spesifieke kombinasie van mikrobiese hidrolitiese ensieme te identifiseer wat die maserasie van rooibos plantmateriaal sal verhoog met behoud van die anti-oksidant inhoud. Tydens hierdie studie is 11 swamspesies wat bekend is vir die produksie van hidrolitiese ensieme, asook 12 swamspecies wat vanaf rooibostee produkte geïsoleer is, geëvalueer vir hul potensiaalom geurontwikkeling en/of ekstraksie van oplosbare stowwe en/of anti-oksidante vanuit rooibostee materiaal te verbeter. Die kru ensiemekstrakte van die 23 isolate, wat ID Aartappel-Dextrose medium opgegroei is, is op oorskot rooibostee geëvalueer vir verhoogde ekstraksie van oplosbare vastestowwe (SS) en/of totale polifenole (TP). Nege rasse het die opbrengs van oplosbare vastestowwe verhoog (verbetering tussen 3% en 42%), terwyl 14 rasse die totale polifenoliese vlakke laat toeneem het (tot so hoog as 36%). Baie min verbetering in kleurontwikkeling van groen (ongefermenteerde) rooibostee is waargeneem, maar ensiemekstrakte van Pleurotus ostreatus var. florida, Lentinula edodes, Aspergillus oryzae, Aspergillus tubingensis, Paecilomyces variotti en Trichoderma reesei, het wel die aroma ontwikkeling vanaf groen tee tot 'n mate verbeter. Tienvoudig gekonsentereerde ekstrakte van vier van hierdie isolate het 'n verbetering van meer as 10% in die ekstraksie van opgeloste vastestowwe uit groen tee tot gevolg gehad. Die ensiemekstrakte van drie swarnme bekend vir hul gebruik in die voedselindustrie, nl. A. oryzae, L. edodes and P. ostreatus var. florida, het relatief hoë vlakke van endoglukanase, xylanase en pektinase aktiwiteit getoon. Agt verskillende kultuur-media is vir die optimale produksie van hidrolitiese and lakkase ensieme vanaf hierdie voedsel-graad swarnme geëvalueer. MYPG was die beste groeimedium vir L. edodes, -terwyl 1% koringstrooi, 1% oorskot graan en 1% pynappelskil die beste induksie van xylanase, pektinase, endoglukanase en lakkase aktiwiteite vir hierdie organisme getoon het. Lentinula edodes opgegroei in YP medium + 1% koringstrooi, het die beste verbetering in aroma en kleur getoon vanaf groen tee getoon. Hierdie ekstrak kan dus moontlik gebruik word vir die verkorting van die fermentasietyd wat vir groen tee benodig word. Ope-lug fermentasie van groen tee duur gewoonlik tot 16 uur en lei tot 'n aansienlike verlies in antioksidant-inhoud. Die R. oryzae ekstrak het die beste potensiaal vir die vervaardiging van 'n "quick-draw" tee getoon met 'n goeie kleurvrystelling sonder enige verlies in SS en TP opbrengs. Wanneer die swamme in Aartappel-Dextrose medium opggegroei word, kan die L. edodes ensiemekstrak vir aroma en kleurontwikkeling van groen tee aangewend word, terwyl die R. oryzae ensiemekstrak vir die verhoging van die antioksidant-inhoud in rooibos ekstrakte van groen tee of gefermenteerde tee gebruik kan word. Dit is bevestig met die kleinskaalse behandeling van gefermenteerde tee waar die L. edodes YP + 1% koringstrooi ensiemekstrak die vrystelling van SS met meer as 30% en die TP met meer as 20% verbeter het.

Thesis (MSc Food Sc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Application of steam treatment to green Cyclopia maculata and C. longifolia to modulate their aroma and flavour profile, without detrimental effects on colour and individual phenolic content, was investigated. Steam treatment (96 °C, atmospheric pressure) of different time periods was applied to the shredded, fresh plant material before drying (STBD; 0, 30, 60, 90 and 120 s) or the herbal tea product after drying (STAD; 0, 1, 2, 3, and 4 min). Steam treatment of 60 s or longer resulted in a significant (p < 0.05) decrease in vegetative (‘green grass’ and ‘hay/dried grass’) and cereal (‘oats/porridge/grains’) aroma and flavour intensities, with an increase in some fruity (‘tropical fruit’ and ‘guava’) and ‘fruity-sweet' aroma intensities. These changes manifested to a greater degree for plant material subjected to STBD, while they were less prominent for C. longifolia compared to C. maculata. Additionally, 19 aroma compounds were instrumentally identified in the volatile fraction of C. maculata infusions prepared from STBD plant material. Of these, seven correlated well with green honeybush vegetative and cereal aroma attributes associated with unsteamed samples. Steaming thus resulted in the rapid loss of some of these highly volatile ‘green’-associated aroma compounds. Steam treatment of more than 60 s, regardless of STBD or STAD, was not detrimental to green colour, and individual phenolic content was not greatly affected. The 30 s STBD treatment of C. maculata, however, led to the rapid loss of green colour and oxidation of phenolic compounds, without considerable aroma improvement. This highlights the need for process control during green honeybush production. Storage stability of steamed (60 s, STBD) and unsteamed green C. maculata was investigated under low (0 °C; moisture impermeable packaging) and normal (25 °C at 60% relative humidity (RH); semi-moisture-permeable sachets) temperature storage conditions (LTS and NTS, respectively) over 6 months and high temperature storage (HTS) conditions (40 °C at 75% RH; semimoisture- permeable sachets) for 1 month. HTS conditions, after 1 month, seemed to emulate changes occurring over 6 months at NTS conditions, with little change detected at LTS conditions. Despite noticeable green colour loss over the respective storage periods at NTS and HTS conditions, individual phenolic compounds were not severely affected. NTS and HTS led to the progressive development of sought-after fermented honeybush sensory attributes, especially in unsteamed samples. These include prominent fruity (‘stewed fruit’, ‘apricot jam’ and ‘marmalade’) aromas and flavours, ‘general sweet’ and ‘fruity-sweet’ aromas and a sweeter taste. The collated descriptive sensory data were used to generate a preliminary sensory wheel for green honeybush aroma, and another for flavour, taste and mouthfeel. The sensory profile of green honeybush was finally described as a dominant vegetative aroma and flavour, prominent sweetassociated and slightly fruity aroma, with sweet and notably bitter tastes and an astringent mouthfeel. By combining the sensory data, it was clear that the major effects of storage outweighed those of steam treatments, suggesting that storage of 3 to 6 months may improve sensory quality, although colour may be slightly compromised. Immediate sensory manipulation and thus improvement, however, may be achieved to a lesser degree in the short term by STBD. / AFRIKAANSE OPSOMMING: Die toepassing van stoombehandeling van groen Cyclopia maculata en C. longifolia vir die modulasie van hul aroma en geur profiel, sonder gepaardgaande nadelige gevolge op kleur en individuele fenoliese inhoud, is ondersoek. Stoombehandeling (96 °C, atmosferiese druk) van verskeie tydstippe is toegepas op die gekerfde, vars plantmateriaal voor droging (SBVD; 0, 30, 60, 90 en 120 s) of die groen teeproduk ná droging (SBND; 0, 1, 2, 3 en 4 min). Stoombehandeling van 60 s of langer het tot 'n betekenisvolle (p < 0.05) afname in vegetatiewe (‘groen gras’ en ‘hooi/gedroogde gras’) en graan (‘hawer/pap/graan’) aroma en geur intensiteite gelei, met 'n toename in sommige vrugtige (‘tropiese vrugte’ en ‘koejawel’) en ‘vrugtige-soet’ aroma intensiteite. Hierdie veranderinge was meer merkbaar in die SBVD behandeling, terwyl C. longifolia minder vatbaar vir hierdie veranderinge was as C. maculata. Daarbenewens is 19 aromaverbindings in die vlugtige fraksie van C. maculata aftreksel voorberei van SBVD plantmateriaal instrumenteel geïdentifiseer. Sewe hiervan het goed gekorreleer met heuningbos vegetatiewe en graan aromas wat verband hou met ongestoomde monsters. Stoombehandeling het dus gelei tot die vinnige verlies van sommige van hierdie baie vlugtige ‘groen’-geassosieërde aromaverbindings. Stoombehandeling vir langer as 60 s, ongeag van SBVD of SBND, het nie die groen kleur en individuele fenoliese inhoud grootliks nadelig beïnvloed nie. ‘n SBVD behandeling van 30 s het egter tot die vinnige verlies van groen kleur en oksidasie van fenoliese verbindings, sonder aansienlike aroma verbetering van C. maculata gelei. Dít beklemtoon die noodsaaklikheid vir prosesbeheer tydens groen heuningbosproduksie. Stabiliteit van gestoomde (60 s, SBVD) en ongestoomde groen C. maculata tydens opberging onder lae (0 °C; vog-ondeurlaatbare verpakking) en normale (25 °C by 60% relatiewe humiditeit (RH); semi-vogdeurlaatbare sakkies) temperatuur opbergingtoestande (LTO en NTO, onderskeidelik) oor 6 maande en hoë temperatuur opbergingtoestande (40 °C by 75% RH; semivogdeurlaatbare sakkies; HTO) vir 1 maand, is ondersoek. HTO het na 1 maand soortgelyke veranderinge teweeggebring as NTO na 6 maande, terwyl min verandering by LTO bespeur is. Ten spyte van merkbare groen kleur verlies oor die onderskeie tydperke by NTO en HTO is individuele fenoliese verbindings min beinvloed. NTO en HTO het gelei tot die progresiewe ontwikkeling van gesogte sensoriese eienskappe van gefermenteerde heuningbos, veral in ongestoomde monsters. Dit sluit in prominente vrugtige (‘gestoofde vrugte’, ‘appelkooskonfyt’ en ‘marmelade’) aromas en geure , ‘algemene soet’ en ‘vrugtige-soet’ aromas en 'n soeter smaak. Al die beskrywende sensoriese data is saamgestel om 'n voorlopige sensoriese wiel vir groen heuningbos aroma op te stel, asook een vir geur, smaak en mondgevoel. Die sensoriese profiel van groen heuningbos kan beskryf word as 'n oorheersende vegetatiewe aroma en geur, prominente soet-verwante en effens vrugtige aroma, met soet en veral bitter smaak en ‘n vrank mondgevoel. Deur die sensoriese data te kombineer kon duidelik uitgewys word dat die belangrikste gevolge van opberging dié van stoombehandeling oortref. Dit dui daarop dat opberging van 3 tot 6 maande sensoriese kwaliteit kan verbeter, hoewel kleur effens benadeel kan word. In die kort termyn kan onmiddellike sensoriese manipulasie en dus verbetering egter in 'n mindere mate deur SBVD bereik word.

The process of classification of the raw material, is one of the most important procedures in any tea dryer, being responsible for ensuring a good quality of the final product. Currently, this process in most tea processing companies is usually handled by an expert, who performs the work manually and at his own discretion, which has a number of associated drawbacks. In this work, a solution is proposed that includes the planting, design, development and testing of a prototype that is able to correctly classify photographs corresponding to samples of raw material arrived at a dryer, using intelligence techniques (IA) type supervised for Classification by Artificial Neural Networks and not supervised with K-means Grouping for class preparation. The prototype performed well and is a reliable tool for classifying the raw material slammed into tea dryers.

La majorité des études sur le thé au Japon se consacre à la cérémonie du thé. Dans cette étude, nous nous proposons au contraire d’analyser l’histoire du thé au Japon du strict point de vue des techniques de culture et de fabrication du thé à l’époque Edo (1600-1868). Dans un premier temps, l’étude se concentre sur l’histoire du thé au Japon depuis son apparition au VIIIe siècle jusqu’au milieu du XIXe siècle. Cette partie abordera l’ensemble du développement de la filière du thé en s’attachant aux points suivants : l’évolution de la consommation du thé sur l’archipel, les différentes techniques de fabrication utilisées et enfin la mise en place des circuits économiques nécessaires au commerce intérieur du thé. Cette étude tentera de comprendre le passage d’une consommation par les élites à une consommation par l’ensemble de la population, ainsi que les implications de cette évolution. Dans un second temps, ce travail se focalisera sur l’étude des techniques de culture et de fabrication du thé à l’époque Edo. De multiples sources historiques sont examinées pour cela : comptes rendus de voyages de savants occidentaux, documents iconographiques divers, mais ce travail s’attache plus spécifiquement à l’analyse des nombreux traités agricoles de l’époque Edo. Ainsi, la description détaillée des différents procédés de fabrication de cette époque permet de comprendre les liens qu’il peut exister entre les différentes techniques, et de réfléchir aux modalités de création et d’amélioration des thés japonais. / Most studies related to Japan’s tea are about the tea ceremony. On the contrary, in this study we are proposing to analyze the history of tea in Japan from the strict point of view of tea culture and manufacture’s techniques in the Edo period (1600-1868). First of all, the study focuses on the history of Japan’s tea from its emergence in the 8th century, up until the mid-19th century. This part will deal with the whole development of the tea sector, focusing on the following points: the evolution of tea consumption on the archipelago, the different manufacturing techniques used and finally the implementation of the economic systems, necessary to the domestic trade. This study is trying to understand the transition from the consumption by the elites only, to the consumption by the entire population, as well as the implications of this evolution. Finally, this study will focus on the study of tea’s culture and manufacture techniques during the Edo period. To do so, we will look at many historical sources: reports and diaries of Western travelers, various iconographic documents, but this thesis will mainly be focusing on the analysis of many agricultural treaties during the Edo period. Therefore, this detailed description of the various manufacturing processes of this period makes it possible to understand the existing links between the different techniques, and to think about the methods of creation and improvement of Japanese teas.