Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXI
Written by Ratu SAFITRI, Mia MIRANTI, Ismi Dwi ASTUTI, Lien KARTIAWATI, Jetty NURHAYATI

Immobilization of cells is a method for placing the microbial cells on a carrier material, wherein the catalytic activity of the cells is still high after storage for a long periode of time. The purpose of this study was to determine the optimum formula for the carrier material inoculum for bioethanol fermentation. The carrier material tested were based on different combination of wheat flour: rice flour, tapioca starch: corn starch and alginate. The study consisted of three phases: First phase is the preparation of the encapsulation of Zymomonas mobilis, Kluyveromyces marxianus, and Saccharomyces cerevisiae. The Second phase is the enzyme hydrolysis process and the third is fermentation of the hydrolysate into bioethanol by the consortium on the best carrier. The method used in this research is descriptive and experimental. Descriptive method carried out in the second phase. While the experimental method with completely randomized design is used in the research phase I and III. The consortium used were KI (Z.mobilis and S.cerevisiae), K2 (Z. mobilis and K marxianus), and K3 (Z. mobilis, S.cerevisiae and K. marxianus). The data were statistically analyzed by ANOVA, and followed by Duncan Test in 5% significance level. The results showed that S.cerevisiae and K.marxianus on formula with a combination of wheat flour: rice flour is the best carrier material with cell density about 3,719x1010 CFU / ml and 3,027x1010 CFU / ml. Meanwhile, alginate is the best carrier for Z.mobilis with cell density about 3,576x1010 CFU / ml. Best consortium in the process of bioethanol fermentation from cellulose waste is Z.mobilis and S.cerevisiae (K1), which have the highest ethanol concentration about 7.167%, the efficiency of fermentation about 61.2%, specific growth rate of 0,072 cells / hour, ethanol yield (Yp/s) 0,23 g/g, microbial yield (Yx/s) 0,33 g/g and a maximum ethanol productivity (qp) of 0,58 g/g/h.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXI
Written by Cătălin STOIAN, Oana LIVADARIU, Mihaela TURTURICĂ, Nicoleta STANCIUC, Liliana MIHALCEA

This paper aims to assess the viability of usingn novel oil extraction methods for Sea Buckthorn (Hippophae rhamnoides L.). Supercritical fluid extraction (SCFE) although extensively used for oil extraction in other countries, is not commercially used in Romania at the moment. Cost constraints, as well as the ease of us, more established methods such as solvent extraction and cold pressing have delayed the implementation of such technologies. Three oil sources were investigated: oils extracted from dry berries using SCFE and cold pressing, and oil extracted from draff (residues after juice extraction) using SCFE. The oils have been analyzed using a HPLC unit, and their carotenoid levels were compared. The results show a slight variation in the carotenoid composition in relation to the extraction methods. This suggests that the SCFE method is viable to be used for large scale Sea Buckthorn oil production.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XX
Written by Ramona Iuliana SZASZ-ZIMA, Monica ENACHE

Honey is a natural, very complex product that can be adulterated using different methods. A reliable determination of honey quality includes complex physico-chemical analyses, organoleptic evaluation as well as microscopic identification of the pollengrains. From the large offer of different honey types made in Romania that are commercially available today, ten brands of monofloral honey bought in Bucharest were analyzed in our lab and the results were compared to the honey standards and other relevant information found in the literature. Simple indicators of quality such as fragrance, appearance and consistency, and the identification of the most frequently occurring pollen helped estimate the accurate labeling of the product. The present study includes the results of the analysis for the samples of 4 monofloral honey types, namely acacia honey, lime honey, oilseed rape honey and raspberry honey.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXI
Written by Adina-Alexandra BAICU, Mona Elena POPA, Elisabeta Elena POPA, Amalia Carmen MITELUŢ

Together with the growth and development of the European Union, the European Food Law has experienced fundamental modifications. If before the Bovine Spongiform Encephalopathy scandal, the food safety legislation was market oriented, nowadays its direction converges to the protection of the consumer. As a result, the actual food safety set of laws bases itself on scientific opinion, together with other factors that the risk manager has to consider when legislating. This paper is depicting the steps taken by the European food legislation until reaching its actual form. It starts with describing the frame in which the European Food Law was approached before its changes, and it continues with presenting the drafts needed to build its present shape. In addition, because the current legislation is founded on the concept of risk analysis, the term will be elaborated both from a general perspective but also with respect to its applications in the food industry field.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXII
Written by Hülya GÜL, Semra Gül TEKELİ

In the present study, the effects of dried and ground Rosa damascena Mill. petals (Rosa flour-RF) for improving the nutritional and some characteristics of gluten-free cookies (GFC) were investigated. The RF was used to replace 0-2.5- 5-7.5-10% of gluten free flour (GFF) formulations. RF supplementation increased lightness and yellowness but decreased redness whereas a slight increase was observed at both moistures and spread ratio. RF addition lead to a significant reduction in the hardness of GFC while different levels of RF did not shown such an effect. RF provided darker samples and cookies became more brittle. RF addition led to an increase in total dietary fibre, total phenolics, and antioxidant activity of GFC. GFC containing RF were more appreciated than GFC with no added RF regarding taste and aroma. At the same time, a higher purchasing decision was found for cookies with added RF. Results showed that acceptable GFC couldbe produced by using RF up to 7.5% level to enhance the nutritional value of GFC that could be important for the gluten-free industry.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XX
Written by Andreea DOBRE, Laura Alexandra MARIN, Carmen MANOLE, Nicoleta ANDREI, Călina Petruța CORNEA

Microorganisms are the main responsible for bioremediation processes. They have the capacity to convert toxic substances in lesser or untoxic ones and also can solubilize compounds in order to make them available to plants. This article emphasizes the capacity of several microorganisms, isolated from different sources, to solubilize Pin, Porg and Zn based compounds. The screening methods comprised the use of several culture media (PVK, NBRIP, PSM and Bunt&Rovira) with added dyes used as pH markers (bromphenol blue and bromcresol purple). The used bacteria were three strains of Bacillus amyloliquefaciens (BIR, BW, OS15, OS17), four strains of Bacillus subtilis (BPA, ICPA, ATCC6633, 10), two strains of Bacillus spp. (B3, B4), one strain of Kluyvera ascorbata(35) and one unindentified strain isolated from soil contaminated with swines manure. After 1 to 7 days of incubation, the halos formed around the colonies showed the bacteria capacity to degrade the compounds of interest. Depending on the screening assay it was determined the eventual mechanisms involved, such as organic acids production, phytase or phosphatases synthesis, etc. Bacteria which had the capacity to solublize Pin based compounds were B3, B4, OS15, BPA, BIR, BW, OS17, ATCC6633, 35 and 10. The ones which were able to degrade phytate (Porg) were BIR, BPA, ICPC, OS15, OS17, BW, 10 and 35. The strains that solublized ZnO were BPA, B4, ATCC 6633, 32 and 35. The isolation sources of bacteria were mainly different types of soil, fact that strengthens the teory that soil bacteria are able to secret metabolites which could be involved in bioremediation and crop protection because they are already adapted to soil contaminants from environment.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXI
Written by Gabriela POPA, Matilda CIUCĂ, Călina Petruța CORNEA, Bogdan NICOLCIOIU, Radu TOMA

Pleurotus ostreatus is one of the most widely cultivated and popular edible fungi from the genus Pleurotus. In the last years, genetic variation of wild strains and cultivated varieties was investigated by use molecular markers and revealed a reduction of the diversity in commercial cultures. In this paper are presented the results obtained by application of combinatorial tubulin based polymorphism (cTBP) and inter simple sequence repeat (ISSR) markers, in order to investigate the genetic variability in several commercial strains of P. ostreatus collected from different geographical locations and their genetic relationships with wild isolates. Five polymorphic PCR products were obtained with primers designated for intron 1 of beta-tubulin gene, and 12 polymorphic amplicons were detected when primers for intron 2 of the same gene were used. Molecular analysis performed with ISSR primers displayed a polymorphism at intra specific level. The data achieved with the cTBP and ISSR markers revealed a reduced genetic differentiation among the commercial lines dispersed in Europe. However, comparing with commercial lines originated from USA, differences at molecular level were observed, suggesting a possible influence of environmental conditions. Regarding the wild strain of P. ostreatus, the molecular data indicated similarity with all European commercial lines. The results demonstrated that the applied molecular methods could be considered efficient tools to evaluate the genetic diversity in Pleurotus ostreatus.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXI
Written by Iuliana RĂUT, Florin OANCEA, Ana Belén López Santísima TRINIDAD, Mariana CĂLIN, Diana CONSTANTINESCU-ARUXANDEI, Mihaela BADEA DONI, Melania- Liliana ARSENE, Gelu VASILESCU, Tatiana Eugenia ŞESAN, Luiza JECU

The genus Phytophthora causes great damages to agricultural production, especially to potatoes and tomatoes cultures. To face these losses, it is of interest to reduce or inhibit the activity of this aggressive pathogen. Some species of Trichoderma have great potential for the biological control of several plant pathogens, including diseases caused by Phytophthora parasitica, Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, Sclerotinia sclerotiorum etc. The purpose of this study was to assess the biocontrol efficacy of three Trichoderma strains (T. asperellum T36, T. asperellum T50, T. harzianum T78) against Phytophthora parasitica. In vitro tests were carried out using dual culture technique. In vivo tests were carried out with pepper seedlings (Capsicum annuum cv. Lamuyo) and conidial standard suspension of Trichoderma isolates as biocontrol agents. Of the three Trichoderma isolates tested for their effectiveness against mycelial growth of pathogen, T. asperellum T36 (81.2%) exhibited maximum inhibition of P. parasitica, compared with the control, followed by T. asperellum T50 (79.6%) and T. harzianum T78 (77.7%). Likewise for the in vivo test, the inoculation of the Trichoderma biocontrol agent showed that the percentage of dead plants seedlings was significant reduced. T. asperellum T36 is a useful biological alternative to pesticides for the control of P. parasitica in pepper seedlings.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XX
Written by Roxana-Mădălina STOICA, Mișu MOSCOVICI, Caterina TOMULESCU, Narcisa BĂBEANU

Bioactive natural products are a main source of new drugs, functional foods and food additives. Chilli pepper is a very important plant used worldwide as a vegetable, a spice and an external medicine. Capsaicin, the pungent principle of Capsicum sp. (69%) is one of the best-known natural compounds. The extraction of the capsacinoids can be made in many ways, with different types of organic solvents, but the yield varies with peppers variety and the conditions of extraction processing. The use of supercritical fluids, especially carbon dioxide, in the extraction of oleoresins, has increased during the last two decades, because it is a simple, inexpensive, fast, effective and solvent-free sample pretreatment technique in comparison with traditional methods (maceration, Soxhlet). Also, the level of capsaicinoids in the Capsicum fruits can be quantified by organoleptic, spectrophotometry, thin-layer chromatography, gas chromatography and high-performance liquid chromatography methods. Of these, high-pressure liquid chromatography (HPLC) is considered the most reliable and rapid method available for the identification and quantification of capsaicinoids.

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Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XX
Written by Mustafa GERMEC, Irfan TURHAN, Ercan YATMAZ, Nedim TETIK, Mustafa KARHAN

Value-added products such as biofuels, enzymes, polysaccharides, pharmaceuticals, and organic acids can be produced from renewable resources such as carob pods, wheat straw, rice straw, rice husk, sugarcane bagasse, etc. by fermentation. Tea processing waste (TPW) is one of the renewable resources, which contains 13.60% cellulose, 32.16% hemicellulose, and 33.38% lignin. Due to its high carbohydrate content (total 45.76% wt), TPW can be used for production of value-added products. Therefore, the main objectives of this study were undertaken not only to determine the chemical composition of acid-pretreated tea processing waste hydrolysate (APTPWH) but also to investigate the effect of pH, inoculum size, agitation, and nitrogen sources on ethanol production from APTPWH without detoxification by using Saccharomyces cerevisiae in a stirred tank bioreactor. Results showed that it contains no HMF and lactic acid, 16.03 g/L fermentable sugar (FS), 0.43 g/L acetic acid, 2.61 g/L glucose, 4.14 g/L fructose+xylose, and 2.27 g/L phenolics. For batch ethanol fermentations, optimum conditions were found to be pH controlled at 5.5, 5% inoculum size (v/v), 150 rpm agitation speed, and yeast extract, which achieved as 25.55% yield (YP/S), 1.75 g/L ethanol production (P), and 0.38 g/L/h productivity (QP). Consequently, ethanol could be relatively produced from TPW without detoxification.

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