Recent advances in biotechnological production of 2-phenylethanol

2-Phenylethanol (2-PE) is an important aromatic alcohol with a rose-like fragrance. It has been widely applied in the cosmetic, perfume, and food industries and is mainly produced by chemical synthesis. An alternative method for the production of natural flavors and fragrances is the microbial transformation process, which is attracting increasing attention because it is an environmentally friendly process and the products are considered “natural”. The production of 2-PE from L-phenylalanine by biotransformation is possible through the Ehrlich pathway and considerable progress has been made in the development of this process. The present report reviews recent advances in biotechnological production of 2-PE, with emphasis on the strategies used to increase production and the applications of in situ product removal techniques. Future research should focus on product scale-up and product recovery processes for the industrialization of microbial processes.     

Optimization of multienzyme production by two mixed strains in solid-state fermentation

F₃ and F₄ strains of Aspergillus niger were screened from five strains of fungi to produce multienzyme preparations (containing cellulase, hemicellulase, glucoamylase, pectinase, and acidic proteinase) as dietary supplementation. Enzyme activities indicated that 1:4 (F₃ to F₄) was the optimum mixture proportion, and 0.3% (W/W) was the preferable pitching rate. In bran mash containing 54.5% (W/W) water, F₃ and F₄ could produce the supplementation better when cultured 30 to 36 h at 30 °C. Monofactorial and orthogonal experiments were performed to optimize media. Results of the variance and range analysis showed that the optimum medium contained 80 g of bran, 20 g of cottonseed powder, 1 g of (NH₄)₂SO₄, and 0.1 g of KH₂PO₄. When F₃ and F₄ strains were cultured in the optimum medium containing 54.5% (W/W) water, the activity of cellulase, hemicellulase, glucoamylase, pectinase, and acidic proteinase reached 996; 15,863; 13,378; 7,621; and 5,583 U/g, respectively.     

Bioproduction of 2-phenylethanol and 2-phenethyl acetate by Kluyveromyces marxianus through the solid-state fermentation of sugarcane bagasse

Applied Microbiology and Biotechnology - 2-Phenylethanol (2-PE) and 2-phenethyl acetate (2-PEA) are important aroma compounds widely used in food and cosmetic industries due to their rose-like...     

Mixture design as first step for improved glutaminase production in solid-state fermentation by isolated Bacillus

Aim: Investigation of mixture‐design impact on glutaminase production by isolated Bacillus sp. Methods and Results: An augmented simplex centroid design was used to optimize a three (wheat bran, Bengal gram husk and palm seed fibre) component mixture for glutaminase production. Selected substrate materials showed impact on glutaminase production values at individual level by Bengal gram husk [2789 U gds^?1 (gram dry substrate] and in two‐level combination with wheat bran and Bengal gram husk (maximum of 3300 U gds^?1). Conclusion: Bengal gram husk is the most suitable substrate medium for glutaminase production by Bacillus sp. Maximum glutaminase production is achieved using solid‐substrate mixture at two‐level combinations in the ratio of 66 : 34 for Bengal gram husk and wheat bran, respectively. Significance and Impact of the Study: The present study has significance in large‐scale production of glutaminase at commercial level with the use of multi‐substrate rather than single‐substrate/support material.     

生物转化法合成2-苯乙醇的研究进展

2-苯乙醇(2-phenylethanol,2-PE)是具有玫瑰香味的芳香醇,在食品、化妆品以及药品领域有着广泛的应用。但是从植物花卉中提取的天然2-苯乙醇产量低,成本高。目前,用微生物转化生产"天然"2-苯乙醇越来越受到关注。本文对近年来国内外报道的提高微生物转化合成2-苯乙醇产量的研究,尤其是生产菌株选育和发酵工程优化的研究进行了综述,并提出今后研究的新思路,旨在为利用微生物发酵进行2-苯乙醇生产提供参考。     

De-hairing protease production by an isolated Bacillus cereus strain AT under solid-state fermentation using cow dung: Biosynthesis and properties

Agro-industrial residues and cow dung were used as the substrate for the production of alkaline protease by Bacillus cereus strain AT. The bacterial strain Bacillus cereus strain AT produced a high level of protease using cow dung substrate (4813 ± 62 U g⁻¹). Physiological fermentation factors such as the incubation time (72 h), the pH (9), the moisture content (120%), and the inoculum level (6%) played a vital role in the enzyme bioprocess. The enzyme production improved with the supplementation of maltose and yeast extract as carbon and nitrogen sources, respectively. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and zymogram analysis of the purified protease indicated an estimated molecular mass of 46 kDa. The protease enzyme was stable over a temperature range of 40–50 °C and pH 6–9, with maximum activity at 50 °C and pH 8. Among the divalent ions tested, Ca²⁺, Na⁺ and Mg²⁺ showed activities of 107 ± 0.7%, 103.5 ± 1.3%, and 104.6 ± 0.9, respectively. The enzyme showed stability in the presence of surfactants such as sodium dodecyl sulfate and on various commercially available detergents. The crude enzyme effectively de-haired goat hides within 18 h of incubation at 30 °C. The enzymatic properties of this protease suggest its suitable application as an additive in detergent formulation and also in leather processing. Based on the laboratory results, the use of cow dung for producing and extracting enzyme is not cumbersome and is easy to scale up. Considering its cheap cost and availability, cow dung is an ideal substrate for enzyme bioprocess in an industrial point of view.     

Oxytetracycline production by Streptomyces rimosus in solid-state fermentation of corncob

Corn-cob was used as a substrate in the production of oxytetracycline by Streptomyces rimosus TM-55 in a solidstate fermentation. Oxytetracycline was detected on day 4, and reached its maximum on day 8. Optimal conditions for oxytetracycline production were an initial pH of 5.2 to 6.3, an initial moisture content of 64% to 67%, supplementation with 20% (w/w) rice bran or 1.5% to 2.5% (w/w) (NH₄)₂SO₄ as sole N source, 1.0% (w/w) CaCO₃, 2% (w/w) MgSO₄.7H₂O, and 0.5% (w/w) KH₂PO₄, with incubation for 8 days at 25 to 30°C. Each g substrate produced 7 to 8 mg oxytetracycline.     

微生物发酵产二十二碳六烯酸代谢机理的研究进展

二十二碳六烯酸(简称DHA)是一种重要的长链多不饱和脂肪酸,对人体具有重要的生理功能。微生物发酵生产的DHA与鱼油来源的DHA相比,具有诸多优点,其发展前景广阔。以下从发酵菌株、代谢途径、关键酶以及油脂积累机制等方面进行了综述,为通过代谢工程等技术手段进一步提高DHA产量提供了参考。     

Modeling of an integrated fermentation/membrane extraction process for the production of 2-phenylethanol and 2-phenylethylacetate

An unstructured model for an integrated fermentation/membrane extraction process for the production of the aroma compounds 2-phenylethanol and 2-phenylethylacetate by Kluyveromyces marxianus CBS 600 was developed. The extent to which this model, based only on data from the conventional fermentation and separation processes, provided an estimation of the integrated process was evaluated. The effect of product inhibition on specific growth rate and on biomass yield by both aroma compounds was approximated by multivariate regression. Simulations of the respective submodels for fermentation and the separation process matched well with experimental results. With respect to the in situ product removal (ISPR) process, the effect of reduced product inhibition due to product removal on specific growth rate and biomass yield was predicted adequately by the model simulations. Overall product yields were increased considerably in this process (4.0 g/L 2-PE+2-PEA vs. 1.4 g/L in conventional fermentation) and were even higher than predicted by the model. To describe the effect of product concentration on product formation itself, the model was extended using results from the conventional and the ISPR process, thus agreement between model and experimental data improved notably. Therefore, this model can be a useful tool for the development and optimization of an efficient integrated bioprocess.     

Medium optimization for the production of the aroma compound 2-phenylethanol using a genetic algorithm

Using a genetic algorithm, 13 medium constituents and the temperature were varied to improve the bioconversion of -phenylalanine ( -phe) to 2-phenylethanol (2-PE) with Kluyveromyces marxianus CBS 600. Within four generations plus an additional temperature screening, corresponding to 98 parallel experiments altogether, a maximum 2-PE concentration of 5.6 g/l, equivalent to an increase of 87% compared to the non-optimized medium was obtained. The vitamin content of the medium had to be raised significantly.     

Production of polyhydroxyalkanoates (PHAs) from waste materials and by-products by submerged and solid-state fermentation

Polyhydroxyalkanoates are biodegradable polymers produced by prokaryotic organisms from renewable resources. The production of PHAs by submerged fermentation processes has been intensively studied over the last 30 years. In recent years, alternative strategies have been proposed, such as the use of solid-state fermentation or the production of PHAs in transgenic plants. This paper gives an overview of submerged and solid-state fermentation processes used to produce PHAs from waste materials and by-products. The use of these low-cost raw materials has the potential to reduce PHA production costs, because the raw material costs contribute a significant part of production costs in traditional PHA production processes.     

Screening of yeasts for the production of the aroma compound 2-phenylethanol in a molasses-based medium

Fourteen yeast strains were screened for production of 2-phenylethanol from l-phenylalanine with molasses as carbon source. Up to 1 g 2-phenylethanol l^–1 was obtained. Using oleyl alcohol as a second phase for in situ product removal to enhance the production of 2-phenylethanol increased the yield to about 3 g 2-phenylethanol l^–1 at 35 °C. The most productive strains were Kluyveromyces marxianus CBS 600 and CBS 397.     

Exploitation of biological wastes for the production of value-added products under solid-state fermentation conditions

Biological wastes contain several reusable substances of high value such as soluble sugars and fibre. Direct disposal of such wastes to soil or landfill causes serious environmental problems. Thus, the development of potential value-added processes for these wastes is highly attractive. These biological wastes can be used as support-substrates in solid-state fermentation (SSF) to produce industrially relevant metabolites with great economical advantage. In addition, it is an environmentally friendly method of waste management. This paper reviews the reutilization of biological wastes for the production of value-added products using the SSF technique.     

辅酶Q_(10)产生菌发酵条件的优化

对辅酶Q10生产菌株鞘氨醇单胞菌YZ0803的发酵条件进行优化,确定发酵时间为90 h,250 mL摇瓶装液量为30 mL。培养基组成(质量分数,下同):葡萄糖1.5%,淀粉2.5%,黄豆饼粉2.5%,(NH4)2SO40.5%,NaCl0.03%,K2HPO40.02%,MgSO40.005%。优化后的辅酶Q10产量达到192 mg/L,比采用基础培养基的产量(138mg/L)提高了39.13%。     

Effect of Pb2+ on the production of hydroxyl radical during solid-state fermentation of straw with Phanerochaete chrysosporium

Hydroxyl radical (OH) is a radical species highly destructive for lignin during solid-state fermentation (SSF) of straw with Phanerochaete chrysosporium (Pc). The production of OH at different initial Pb²⁺ concentrations during SSF of straw with Pc was investigated. The results showed that a modest amount (under 200 mg kg⁻¹) of Pb²⁺ could enhance the production of OH, while a higher Pb²⁺ concentration resulted in inhibition. The content of OH reached the peak value at day 12 in the whole tested samples, and the maximal content of OH was obtained at initial Pb²⁺ concentration of 100 mg kg⁻¹. It was also found that the production of OH was connected to enzymatic activity and oxalate content in some degree, in particular, a significant positive correlation was found between oxalate concentration and production of OH.We found that low concentration of Pb²⁺ can promote the degradation of lignin, and the higher initial Pb²⁺ concentration (400 mg kg⁻¹) resulted in inhibition. In addition, it appeared that there was no significant correlation between lignin degradation rate and the production of OH when Pb²⁺ concentration was taken into account.     

The behavior of kinetic parameters in production of pectinase and xylanase by solid-state fermentation

Solid-state fermentation (SSF) is defined as the growth of microbes without a free-flowing aqueous phase. The feasibility of using a citrus peel for producing pectinase and xylanase via the SSF process by Aspergillus niger F3 was evaluated in a 2 kg bioreactor. Different aeration conditions were tested to optimize the pectinase and xylanase production. The best air flow intensity was 1 V kg M (volumetric air flow per kilogram of medium), which allowed a sufficient amount of O2 for the microorganism growth producing 265 U/g and 65 U/g pectinases and xylanases, respectively. A mathematical model was applied to determine the different kinetic parameters related to SSF. The specific growth rate and biomass oxygen yield decreased during fermentation, whereas an increase in the maintenance coefficient for the different employed carbon sources was concurrently observed.     

Sugarcane-pressmud as a novel substrate for production of citric acid by solid-state fermentation

Sugarcane-pressmud, a by-product of cane-sugar manufacture, was used as a substrate for production of citric acid by Aspergillus niger CFTRI 30, in a solid-state fermentation system. Of the 170 g of sugar supplied, 131 g were consumed, with a 79% yield of citric acid over 120 h. Potassium ferrocyanide improved the conversion to about 88% and lowered the fermentation time by 24 h. Enrichment with sugar and NH₄NO₃ was essential to improve productivity. About 174 g citric acid/kg dry sugarcane-pressmud were produced after 120 h in ferrocyanid-treated medium which initially contained 12.5% (w/w) effective sugar and 0.1% (w/w) NH₄NO₃. About 3% (w/w) of the original sugar present in the sugarcane-pressmud was non-utilizable. This is the first report on the potential of sugarcane-pressmud for citric acid production.V.S. Shankaranand and B.K. Lonsane are with the Fermentation Technology and Bioengineering Discipline, Central Food Technological Research Institute, Mysore-570 013, India     

Polyurethane foam as an inert carrier for the production of L(+)-lactic acid by Lactobacillus casei under solid-state fermentation

AIM: Production of L-lactic acid in solid-state fermentation (SSF) using polyurethane foam (PUF) as inert support moistened with cassava bagasse starch hydrolysate. METHODS AND RESULTS: PUF impregnated with cassava bagasse starch hydrolysate as major carbon source was used for the production of L-lactic acid using Lactobacillus casei in solid-state condition. The key parameters such as reducing sugar, inoculum size and nutrient mixture were optimized by statistical approach using response surface methodology. More than 95% conversion of sugars to lactic acid from 4 g reducing sugar per gram dry support was attained after 72 h when the inert substrate was moistened with 6.5 ml of nutrient solution and inoculated with 1.5 x 10(9) CFU of L. casei. While considering the lactate yield based on the solid support used, a very high yield of 3.88 g lactic acid per gram PUF was achieved. CONCLUSION: PUF acted as an excellent inert support for L. casei and provided a platform for the utilization of starchy waste hydrolysate in a lower reactor volume. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a cost effective cultivation of lactic acid bacteria for producing lactic acid from agro based waste products such as cassava bagasse. This is the first report on the exploitation of PUF as an inert support for lactate production under SSF.