Itaconic acid production by immobilizedAspergillus terreus on sucrose medium

Summary The itaconic acid production by immobilizedAspergillus terreus TTK 200-5-3 mycelium was optimized in shake flask fermentations using statistical experimental design and empirical modelling. The maximum itaconic acid concentration was calculated to be 13.3 g/l in the investigated experimental area when initial sucrose concentration was 10%, ammonium nitrate concentration 0.275% and initial pH 3. The itaconic acid product concentration using immobilized mycelium was about double of that obtained with the free mycelium.     

Itaconic acid production from xylose in repeated-batch and continuous bioreactors

Summary Itaconic acid production from xylose by immobilized Aspergillus terreus TKK 200-5-2 mycelia was optimized both in repeated shake-flask fermentations and in continuous column bioreactors using statistical experimental design and empirical modelling. Using continuous 9-1 scale air-lift bioreactors, a pH of 2.5, aeration rate of 0.6 v/v per minute and residence time of 160 h gave the highest itaconic acid concentration. In air-lift bioreactors a cubic carrier size of 0.5 cm gave a 3.3-fold higher product concentration than 1-cm cubes. Packed-bed column reactors had a higher production rate than air-lift reactors. Offprint requests to: H. Kautola     

Itaconic acid production from wheat chaff by Aspergillus terreus

Biotechnologically produced itaconic acid is an important building block for the chemical industry and still based on pure carbon sources, detoxified molasses or starch hydrolysates. Changing these first generation feedstocks to alternative renewable resources of a second generation implies new challenges for the cultivation process of the industrial itaconic acid producer Aspergillus terreus, which is known to be very sensitive towards impurities. To select a suitable pretreatment method of a second generation feedstock, the influences of different hydrolysate components, like monosaccharides and sugar degradation products, were tested. Particular the impact of those components on itaconic acid yield, productivity, titer and morphology was investigated in detail. Wheat chaff was used as lignocellulosic biomass, which is an agricultural residue. An alkaline pretreatment method with sodium hydroxide at room temperature and a subsequent enzymatic saccharification at pH 4.8 at 50 °C with 10 FPU/g_Biomass Biogazyme 2x proved to be very suitable for a subsequent biotechnological production of itaconic acid. A purification by a cation exchanger of the wheat chaff hydrolysate resulted in a final titer of 27.7 g/L itaconic acid with a yield of 0.41 g/g_{total sugar}.     

Itaconic acid production by immobilizedAspergillus terreus from xylose and glucose

Summary Aspergillus terreus NRRC 1960 spores were entrapped in calcium alginate gel beads or alternotely the fungal mycelium was immobilized either on Celite R-626 or in agar gel cubes, and the biocatalyst was employed both in repeated batch and in continuous column reactors to produce itaconic acid from D-xylose or D-glucose. The highest itaconic acid yield obtained in a submerged culture batch fermentation was 54.5% based on total initial glucose (55 g/l) with a volumetric productivity of 0.32 g/l h, and 44.8% from xylose (67 g/l) with a productivity of 0.20 g/l h. In a repeated batch fermentation mycelium immobilized in agar gel had a productivity of 0.112 g/l h, and mycelium grown from spores immobilized in calcium alginate gel 0.06 g/l h, both from xylose (60 g/l). With the best immobilized biocatalyst system used employing Celite R-626 as a carrier, volumetric productivities of 1.2 g/l h from glucose and 0.56 g/l h from xylose (both at 60 g/l) were obtained in continuous column operation for more than 2 weeks.     

Itaconic acid production by Aspergillus terreus on raw starchy materials

Starchy materials such as corn starch, soft wheat flour, potato flour, cassava flour, sorghum starch, sweet potato and industrial potato flours, either acid or enzymatically hydrolysed, were used as substrates for itaconic acid production by Aspergillus terreus NRRL 1960. Both production and yield were highest on corn starch (18·4 g l⁻¹ and 34·0%, respectively). The degree of hydrolysis had a great influence on acid production which was highest when corn starch was saccharified at 85 DE (dextrose equivalent). In a 3 litre benchtop fermenter, itaconic acid production and productivity were 19·8 g l⁻¹ and 0·13 g l⁻¹ h⁻¹, respectively.     

Itaconic acid production by immobilized Aspergillus terreus with varied metal additions

Summary The effect of trace and alkaline metals on itaconic acid production by polyurethane-foam-immobilized Aspergillus terreus was examined in repeated shake-flask cultures according to a statistical experimental design. An increase in the glucose or copper concentration increased the need for earth alkaline metals. The experimentally obtained highest itaconic acid concentration of 51 g/l from 15% glucose with a total productivity of 3.67 g/l per day was reached during the first 14-day batch fermentation. In the fourth batch the calculated highest itaconic acid concentration of 19 g/l was reached with 25% glucose, 5 g/l of magnesium sulphate, 13 mg/l of copper sulphate and 10 g/l of calcium chloride. The immobilization of the mycelium increased the itaconic acid concentration obtained by as much as eightfold.Offprint requests to: H. Kautola     

The use of microorganisms in L-ascorbic acid production

L-Ascorbic acid has been industrially produced for around 70 years. Over the past two decades, several innovative bioconversion systems have been proposed in order to simplify the long time market-dominating Reichstein method, a largely chemical synthesis by which still a considerable part of L-ascorbic acid is produced. Here, we describe the current state of biotechnological alternatives using bacteria, yeasts, and microalgae. We also discuss the potential for direct production of l-ascorbic acid exploiting novel bacterial pathways. The advantages of these novel approaches competing with current chemical and biotechnological processes are outlined.     

Engineering robust microorganisms for organic acid production

Organic acids are an important class of compounds that can be produced by microbial conversion of renewable feedstocks and have huge demands and broad applications in food, chemical, and pharmaceutical industries. An economically viable fermentation process for production of organic acids requires robust microbial cell factories with excellent tolerance to low pH conditions, high concentrations of organic acids, and lignocellulosic inhibitors. In this review, we summarize various strategies to engineer robust microorganisms for organic acid production and highlight their applications in a few recent examples.     

Itaconic acid production using an air-lift bioreactor in repeated batch culture of Aspergillus terreus

Repeated itaconic acid production using an air-lift bioreactor was carried out by three methods—two with cell recycling by means of centrifugation and filtration by a stainless steel filter set inside the bioreactor and one by repeated batch culture without cell recycling. In a flask culture, repeated itaconic acid production was stable for 9 cycles (45 d) and the production rate was 0.47 g/l/h. However, in the air-lift bioreactor, it was difficult to produce itaconic acid in the repeated batch culture with cell recycling for a long period due to a decrease in fluidity resulting from an increase in mycelium concentration. In the method without cell recycling, however, repeated itaconic acid production was stable for 4 cycles (21 d) and the production rate was 0.37 g/l/h.     

工业乳酸发酵的近期进展

乳酸是一种重要的多用途有机酸。通过菌种改良和发酵工艺技术的改进,可以大大提升微生物发酵技术水平,降低成本。简要综述有关的研究进展。     

Glycolic acid production using ethylene glycol-oxidizing microorganisms.

Screening for microorganisms oxidizing ethylene glycol to glycolic acid was carried out. Among stock cultures, several yeasts and acetic acid bacteria showed high glycolic acid producing activity. Pichia naganishii AKU 4267 formed the highest concentration of glycolic acid, 35.3 g/l, from 10% (v/v) ethylene glycol (molar conversion yield, 26.0%). Among soil isolates, Rhodotorula sp. 3Pr-126, isolated using propylene glycol as a sole carbon source, formed the highest concentration of glycolic acid, 25.1 g/l, from 10% (v/v) ethylene glycol (molar conversion yield, 18.5%). Rhodotorula sp. 3Pr-126 showed higher activity toward 20% (v/v) ethylene glycol than P. naganishii AKU 4267. Optimization of the conditions for glycolic acid production was investigated using P. naganishii AKU 4267 and Rhodotorula sp. 3Pr-126. Under the optimized conditions, P. naganishii AKU 4267 and Rhodotorula sp. 3Pr-126 formed 105 and 110 g/l of glycolic acid (corrected molar conversion yields, 88.0 and 92.2%) during 120 h of reaction, respectively.     

Itaconic acid production using sago starch hydrolysate by Aspergillus terreus TN484-M1

Sago starch was hydrolyzed using either chemical agents, or enzymes at various pH and concentrations. Hydrolysis using 5000 AUN/ml (0.5%, w/v) glucoamylase exhibited the highest itaconic acid yield up to 0.36 g/g sago starch, whereas hydrolysis using nitric acid at pH 2.0 yielded 0.35 g/g sago starch. The medium was optimized and the composition was (g/l) 140 sago starch, 1.8 corn steep liquor, 1.2 MgSO(4).7H(2)O and 2.9 NH(4)NO(3). When the optimal conditions of hydrolysis and medium composition were applied to itaconic acid production in a 3-l jar fermentor, the itaconic acid production was 48.2 g/l with a yield of 0.34 g/g sago starch. This was filtered from the cultured broth and 37.1g of itaconic acid was recovered with a purity of 97.2%. This result showed that sago starch could be converted to a value-added product with only a simple pretreatment.     

Towards systems metabolic engineering of microorganisms for amino acid production

Microorganisms capable of efficient production of amino acids have traditionally been developed by random mutation and selection method, which might cause unwanted physiological changes in cellular metabolism. Rational genome-wide metabolic engineering based on systems and synthetic biology tools, which is termed 'systems metabolic engineering', is rising as an alternative to overcome these problems. Recently, several amino acid producers have been successfully developed by systems metabolic engineering, where the metabolic engineering procedures were performed within a systems biology framework, and entire metabolic networks, including complex regulatory circuits, were engineered in an integrated manner. Here we review the current status of systems metabolic engineering successfully applied for developing amino acid producing strains and discuss future prospects.     

Fatty acid biosynthesis in microorganisms being used for Single Cell Oil production

Single cell oils (SCOs) are now produced by various microorganisms as commercial sources of arachidonic acid (ARA) and docosahexaenoic acid (DHA). These oils are now used extensively as dietary supplements in infant formulas. An understanding of the underlying biochemistry and genetics of oil accumulation in such microorganisms is therefore essential if lipid yields are to be improved. Also an understanding of the biosynthetic pathways involved in the production of these polyunsaturated fatty acids (PUFAs) is also highly desirable as a prerequisite to increasing their content in the oils. An account is provided of the biosynthetic machinery that is necessary to achieve oil accumulation in an oleaginous species where it can account for lipid build up in excess of 70% of the cell biomass. Whilst PUFA production in most microorganisms uses a conventional fatty acid synthase (FAS) system followed by a series of desaturases and elongases, in Schizochytrium sp., and probably related thraustochytrid marine protists, PUFA synthesis now appears to be via a polyketide synthase (PKS) route. This route is discussed. It clearly represents a major departure from conventional fatty acid biosynthesis, possibly as a means of decreasing the amount of NADPH that is needed in the overall process.     

Methionine production by microorganisms

Microbial production of methionine is reviewed with 71 references. The review describes different methionine-producing organisms, as well as analog-resistant regulatory mutants, their optimum cultural conditions and yields. The pathways of methionine biosynthesis and their regulation are discussed.     

微生物有机酸转运蛋白的研究进展

转运蛋白是一类膜蛋白,可介导生物膜内外化学物质的跨膜转运及信号交换。有机酸转运蛋白在微生物有机酸代谢的跨膜转运过程中发挥重要作用,根据转运蛋白有机酸转运的方向不同可以分为摄取转运蛋白和外排转运蛋白。在微生物代谢中,有些有机酸可以作为能源直接参与体内代谢,有些是能量转换过程中的重要中间产物;摄取转运蛋白的过表达,可以促进微生物细胞获取能源物质,高效的生产目标产物;有机酸摄取转运蛋白敲除或外排转运蛋白表达,有利于底盘细胞外排更多目标产物,进而促进有机酸的生物合成。研究有机酸转运蛋白的结构和功能,有助于解析微生物细胞有机酸生物合成及利用的机制,对于提高工业微生物对有机酸的利用及生物合成具有重要作用。本文综述了微生物有机酸转运蛋白分类和结构、转运方式和转运功能等方面,重点综述了转运蛋白在有机酸生产中的应用,为工业微生物有机酸的高效生物合成及未来发展提供参考。