Lovastatin production by Aspergillus terreus in a two‐staged feeding operation

BACKGROUND: Lovastatin is known to inhibit its own synthesis in the fungus Aspergillus terreus. Therefore, the use of a fermentation strategy that continuously removes some of the lovastatin produced from the bioreactor can enhance its productivity. This paper reports on the effects of dilution rate and the composition of the feed medium on lovastatin production by A. terreus. RESULTS: The feeding strategy consisted of an initial batch/fed‐batch phase and a semi‐continuous culture phase in which the pelleted biomass was retained inside a slurry bubble column. A nitrogen‐free medium was fed at various fixed dilution rates in the semi‐continuous phase. In experiments that were designed to assess the effects of the composition of the medium, the dilution rate was held at 0.42 d^?1, but different feed media were used in separate runs. The best two‐staged production strategy was shown to consist of a 96 h batch/fed‐batch phase that used a nutritionally complete medium. This was followed by a semi‐continuous operation using a medium that was free of both nitrogen and carbon sources. CONCLUSION: Semi‐continuous operation enhanced productivity of lovastatin by 315% compared with a conventional batch operation. The optimal dilution rate in semi‐continuous operation was about 0.42 d^?1. Copyright © 2008 Society of Chemical Industry     

碳源对土曲霉中洛伐他汀生物合成的影响

通过摇瓶培养考察碳源种类及其初始水平对土曲霉次级代谢物洛伐他汀生物合成的影响。蔗糖和葡萄糖等快速利用碳源产洛伐他汀量较低,且易积累其直接前体monacolin J;淀粉和甘油等缓慢利用碳源有利于细胞生长和产物合成,其monacolin J积累相对较少,甘油为碳源时,可获得最高产物量(937.5±12.3)mg/L,乳糖不利于细胞生长导致较低的产物量。以淀粉为碳源时,其初始水平的增加可促进洛伐他汀的合成,碳质量分数为2%时对底物利用率较高,4%时最高产物量可达(875.4±23.1)mg/L,monacolin J则在2%—3%时积累较多,同时产生较多的2-甲基丁酸。     

Enhanced production of lovastatin in a bubble column by Aspergillus terreus using a two‐stage feeding strategy

A two‐stage feeding strategy is shown to improve the rate of production of lovastatin by Aspergillus terreus when compared with conventional batch fermentation. The feeding strategy consisted of an initial batch/fed‐batch phase and a semi‐continuous culture dilution phase with retention of pelleted biomass in a slurry bubble column reactor. The batch phase served only to build up the biomass for producing lovastatin, a secondary metabolite that inhibits its own synthesis in the producing microfungus. The semi‐continuous dilution phase provided nutrients to sustain the fungus, but prevented biomass growth by limiting the supply of essential nitrogen. (Synthesis of lovastatin does not require nitrogen.) The preferred pelleted growth morphology that favors lovastatin synthesis was readily obtained and maintained in the 20 L bubble column used. In contrast, a stirred tank fermentation had a substantially lower production of lovastatin because mechanical agitation damaged the fungal pellets. The two‐stage feeding method increased lovastatin production rate by more than 50% in comparison with the conventional batch operation. Rheological data for the fungal broth are presented. Copyright © 2007 Society of Chemical Industry     

Fermentation optimization for the production of lovastatin by Aspergillus terreus: use of response surface methodology

A Box–Behnken experimental design was used to investigate the effects of five factors—ie oxygen content in the gas phase; concentrations of C, N and P; and fermentation time—on the concentrations of biomass and lovastatin produced in batch cultures of Aspergillus terreus. The values of the various factors in the experiment ranged widely, as follows: 20–80% (v/v) oxygen in the aeration gas; 8–48 g dm^?3 C‐concentration; 0.2–0.6 g dm^?3 N‐concentration; 0.5–2.5 g dm^?3 phosphate‐concentration; and 7–11 days fermentation time. No previous work has used statistical analysis in documenting the interactions between oxygen supply and nutrient concentrations in lovastatin production. The Box–Behnken design identified the oxygen content in the gas phase as the principal factor influencing the production of lovastatin. Both a limitation and excess of oxygen reduced lovastatin titers. A medium containing 48 g dm^?3 C supplied as lactose, 0.46 g dm^?3 N supplied as soybean meal, and 0.79 g dm^?3 phosphate supplied as KH₂PO₄, was shown to support high titers (～230 mg dm^?3) of lovastatin in a 7‐day fermentation in oxygen‐rich conditions (80% v/v oxygen in the aeration gas). Under these conditions, the culture medium had excess carbon but limiting amounts of nitrogen. The optimized fermentation conditions raised the lovastatin titer by four‐fold compared with the worst‐case scenario within the range of factors investigated. Copyright © 2004 Society of Chemical Industry     

土曲霉M11菌丝球对结晶紫的吸附脱色研究

研究了土曲霉M11菌丝球对水溶液中结晶紫染料的生物吸附作用,考察了菌丝球的死活及培养方式、培养温度、溶液盐度、培养时间、染料溶液初始pH值和接种量对结晶紫脱色率的影响。结果表明,土曲霉M11的菌丝球在35℃1、50 r.min-1振荡培养方式下脱色效果较好,灭活菌丝球脱色效果更好,其24 h内对结晶紫脱色率均达90%以上;培养温度和接种量对菌丝球脱色效果影响不大;在初始pH值为6时,脱色效果最好,对结晶紫脱色率达93%以上。另外,较低的盐浓度对菌丝球脱色效果有一定抑制作用,而较高的盐浓度对结晶紫脱色效果无明显影响。     

5mmminipage15cm A Morphologically Structured Model for Mycelial Growth andSecondary Metabolite Formation

A morphologically structured model is proposed to describe the batch fermentation of lovastatin according to the growth kinetics of filamentous microorganisms. Three kinds of hyphae are considered in the model:actively growing hyphae, non-growing hyphae and deactivated hyphae. Furthermore, actively growing hyphae consist of three morphological compartments: apical compartment which gives rise to hyphal tip extension; subapical compartment which is related to hyphal branching; and hyphal compartment which is only responsible for secondary metabolite formation. The kinetics of mycelial growth mechanism is summarized and applied in modeling lovastatin fermentation. A Michaelis-Menten kinetic model with substrate inhibition is proposed for product formation. As expected, the model simulations fit well with experimental data obtained either from a laboratory scale 10 L fermenter or from a vilot-vlant scale fermenter.     

Fungus-Mediated Green Synthesis of Silver Nanoparticles Using Aspergillus terreus

The biosynthesis of nanoparticles has received increasing attention due to the growing need to develop safe, cost-effective and environmentally friendly technologies for nano-materials synthesis. In this report, silver nanoparticles (AgNPs) were synthesized using a reduction of aqueous Ag(+) ion with the culture supernatants of Aspergillus terreus. The reaction occurred at ambient temperature and in a few hours. The bioreduction of AgNPs was monitored by ultraviolet-visible spectroscopy, and the AgNPs obtained were characterized by transmission electron microscopy and X-ray diffraction. The synthesized AgNPs were polydispersed spherical particles ranging in size from 1 to 20 nm and stabilized in the solution. Reduced nicotinamide adenine dinucleotide (NADH) was found to be an important reducing agent for the biosynthesis, and the formation of AgNPs might be an enzyme-mediated extracellular reaction process. Furthermore, the antimicrobial potential of AgNPs was systematically evaluated. The synthesized AgNPs could efficiently inhibit various pathogenic organisms, including bacteria and fungi. The current research opens a new avenue for the green synthesis of nano-materials.     

Optimized reuse and bioconversion from retentate of pre‐filtered palm oil mill effluent (POME) into microbial protease by Aspergillus terreus using response surface methodology

BACKGROUND: The membrane filtration process enables the treatment of wastewater, producing permeate which is less polluted. However, disposal is usually required for the retentate, which is produced as a concentrated constituent along with the permeate. In this study, the authors explored the possibility of reusing, rather than disposing of, the retentate of pre‐filtered palm oil mill effluent (POME) as a fermentation substrate in protease production by a wild type strain of Aspergillus terreus IMI 282743. In addition, the quantitative and interactive effects of the concentration factor for retentate, temperature, inoculum concentration, and fermentation time on the optimization of protease production were investigated using response surface methodology (RSM). RESULTS: Using RSM, the optimum conditions were found to be a concentration factor of 7.27, temperature of 37.95 °C, inoculum concentration of 1.30% (v/v) and fermentation time of 3.83 days. The protease production was increased 4.37‐fold in comparison with the results obtained under non‐optimized conditions. CONCLUSION: To a certain extent, protease production could be enhanced with an increase in concentration factor and temperature, and a decrease of inoculum concentration and fermentation time. Also, POME retentate was found to be a good substrate for protease production with high product activity and without nutrient supplementation. Copyright © 2009 Society of Chemical Industry     

Bubble column fermentation of olive mill wastewaters by Aspergillus niger

A bubble column (tower) fermenter was used to study the growth of Aspergillus niger on olive mill wastewaters (OMW). The gas hold-up, mixing time and oxygen transfer coefficient values were studied in terms of superficial air velocity. These characteristics were clearly affected by OMW. The growth of A. niger resulted in a filamentous suspension with no pellet formation and the specific growth rate was 0–2 h^?1. The growth rate was limited by the oxygen transfer rate at 90 mmol dm^?3 h^?1 when pO₂ was lower than 2%.     

超声波对铅膏湿法转化产物结晶形貌的影响

通过柠檬酸湿法工艺处理废旧铅酸蓄电池铅膏,浸出转化得到的前体柠檬酸铅结晶产物粒度较小,不易过滤。通过超声波的方式对柠檬酸铅的结晶过程加以控制,对处理前后的样品进行XRD、TG-DTA和SEM表征,对比超声波处理前后的柠檬酸铅晶体形貌变化。结果表明,经过超声波处理后,柠檬酸铅晶体由薄片状转变为长径比约为8∶1的柱状,柱状晶体长20～50 μm,超声波处理前柠檬酸铅晶体的粒度均小于5 μm。因此,超声波处理能够有效调控柠檬酸铅晶体的结晶形貌,改善柠檬酸铅前体的过滤性能,提高铅膏湿法转化的效率。     

Effects of blend composition on the morphologies and physical properties of polycarbonate/acrylonitrile-butadiene-styrene blends

The morphologies and physical properties of twin-screw-extruded polycarbonate/acrylonitrile-butadiene-styrene (PC/ABS) blends with various blend ratios are studied. The needle-like co-continuous phase in PC-rich blends changes to the sea-island phase for blend ratios of more than 50 wt% ABS. While pure PC exhibits an almost-Newtonian flow behavior, PC/ABS blends exhibit the interesting rheological transition. The viscosities of the ABS-rich blends at low shear rates are almost equal to those of the pure ABS polymer. The yield stress for the PC/ABS blend ratio of 3:7 is the highest in composition. At the frequency of 10 rad/s, the PC-rich blends exhibit highly viscous properties, whereas the ABS-rich blends present highly elastic properties as the temperature increases. Moreover, the ABS polymer in the PC/ABS polymer blend induces significant change at the fracture surface of PC, transitioning from brittle to ductile nature.     

复合诱变选育衣康酸高产菌株的研究

采用紫外线-LiCl、硫酸二乙酯(DES)复合诱变,经平板菌落筛选、摇瓶筛选以及连续传代稳定性考察,从土曲霉(Aspergillus terreus)2433出发菌株获得一株衣康酸高产突变株土曲霉At DES-11,其产酸量达到48.22 g/L,较出发菌株提高了156.08%;糖酸转化率为61.82%,较出发菌株提高了101.36%,且连续传代5次后,遗传性状仍较稳定,产酸量稳定在48 g/L左右,菌落及菌丝形态基本不变。     

Effects of chemical modification of wood flour on the rheological properties of high‐density polyethylene blends

Chemical modification of lignocellulosic fibers can improve interfacial adhesion and dimensionally stabilize the resulting plastic composites. This study examined the rheological properties of wood flour/high density polyethylene (HDPE) melts after poplar wood flour was modified with glutaraldehyde (GA, mainly cell wall cross‐linking) and 1,3‐dimethylol‐4,5‐dihydroxyethyleneurea (DMDHEU, mainly poly‐condensation). Results show improvement in both the dispersibility of treated wood flour in the HDPE and its interfacial compatibility. Treatment with GA decreased melt viscosity, moduli, and shear stress as evidenced by rheometry. However, the modifying effects of DMDHEU were not observed, which was mainly due to reduced HDPE content. This study indicates that chemical modification of wood flour is a promising approach to improve the processability of highly filled wood thermoplastic composites via extrusion/injection molding processing. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41200.     

Selection and isolation of glucose-tolerant amylolytic Aspergillus by cyclic fed batch culture process

A simple cyclic fed batch culture system was developed for selection and subsequent isolation of glucose-tolerant Aspergillus niger strain. A culture medium which contained 1.2 g dm⁻³ of glucose was inoculated with a non-glucose-tolerant A. niger (K_i =20.25 g dm⁻³). A culture medium of higher glucose concentration (100 g dm⁻³ and 200 g dm⁻³) was fed at a rate equal to the rate of HN₄⁺ consumption by means of a pH control system. The maximum and minimum liquid levels in the fed batch culture vessel were determined by two liquid level detectors which activated and deactivated a harvest pump. The novelty of the selection system is that the frequency and pressure of selection increase gradually but continuously, and they are determined by the intrinsic potential of the culture. The process was fully automatic. An Aspergillus mutant which had a glucose inhibition constant of 3200 g dm⁻³ was isolated after six generations. The process should be particularly useful for screening filamentous microorganisms growing on novel substrates or tolerating inhibitors.     

AstPT Catalyses both Reverse N1‐ and Regular C2 Prenylation of a Methylated Bisindolyl Benzoquinone

Prenylated bisindolyl benzoquinones exhibit interesting biological activities, such as antidiabetic or anti‐HIV activities. A number of these compounds, including asterriquinones, have been isolated from Aspergillus terreus. In this study, we identified two putative genes by genome mining, ATEG_09980 and ATEG_00702, which share high sequence similarity with the known bisindolyl benzoquinone prenyltransferase TdiB from Aspergillus nidulans. The coding sequences were cloned and overexpressed in E. coli. The overproduced recombinant proteins were purified to near homogeneity and used for enzyme assays with asterriquinone D in the presence of dimethylallyl diphosphate. HPLC analysis showed that product formation was only detected in enzyme assays with EAU29429 encoded by ATEG_09980, not in those with EAU39348 encoded by ATEG_00702. Product isolation and structure elucidation by NMR and MS analyses led to identification of N1‐reversely and C2‐regularly monoprenylated derivatives, as well as N1′,N1′′reversely, N1′‐reversely, C2′′‐regularly diprenylated derivatives. This proved that EAU29429 functions as an asterriquinone prenyltransferase (AstPT) and indicated the involvement of EAU29429 rather than EAU39348 in the biosynthesis of methylated asterriquinones. Furthermore, incubation of monoprenylated enzyme products with AstPT resulted in the formation of the diprenylated derivatives.     

Effects of compatibilizer on the mechanical,rheological, and shape memory behaviors of poly(lactic acid) and poly(MnBM) blends

The effects of compatibilizer on the morphological, mechanical, rheological, and shape memory properties of poly(lactic acid) (PLA) and poly(methyl methacrylate-block-n-butyl acrylate-block-methyl methacrylate) (Poly(MnBM)) (80/20) blend were investigated. From the morphological results, the addition of 1 wt% SAN-MAH as a compatibilizer showed minimum Poly(MnBM) domain size among the blends with the SAN-MAH in the amounts from 0 to 7 wt%. Tensile and flexural strengths, and complex viscosity of the blends showed maximum when the SAN-MAH content was 1 wt%, which suggested the increased compatibility between the PLA and Poly(MnBM) phases. From the above results, the optimum compatibilizer content of the PLA and Poly(MnBM) blend was 1 wt%. The recovery ratio of tensile energy was found to be 83 and 56% for the PLA/Poly(MnBM) blend with and without the SAN-MAH (1 wt%), respectively. Upon blending the PLA and Poly(MnBM) (80/20) with SAN-MAH (1 wt%), the increase of recovered tensile energy was observed, and that the brittleness of PLA was improved to be ductile which resulted an improved shape memory behavior of the blend. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48591.     

Effects of relaxation time and zero shear viscosity on structural evolution of linear low‐density polyethylene in shear flow

The purpose of this work is to investigate the precursor formation and crystallization of four different types of linear low‐density polyethylene in shear flow. The aggregation of string‐like structure (precursor) in micrometer scale was observed by polarized optical microscopy. Although the existence of precursors accelerates crystallization, we find that it is unnecessary for the polymer fluid to possess crystalline structures at relatively high temperatures by wide‐angle X‐ray diffraction. The rotational rheometer result suggests that relaxation time is consistent with processes of the precursor formation observed at 120 °C, while zero shear viscosity affects the boundaries of their corresponding processing windows. According to these results, we propose a mechanism that the precursor formation consists of entanglement and relaxation stages. Entanglements preserve the ordered state of polymer chains, while they return to the initially disordered state during the relaxation stage. Under shearing, the polymer chain is oriented along the flow direction, and the degree of chain motion remains limited because the oriented parts are confined by the entanglements (characterized by zero shear viscosity η₀) acting as slip‐links. However, some chain motions and relaxation (characterized by terminal relaxation time τ) can still take place during this stage. Afterwards, the polymer chain becomes disordered and some entanglements disappear. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46053.     

Inactivation of Aspergillus spp. by Ozone Treatment

The article investigates the effectiveness of ozone in inhibiting the growth of two Aspergillus species, (A. flavus, A. niger) isolated from a nail affected by onychomycosis and from a biodeteriorated paper. Specifically, two main goals are to establish (i) whether differently aged colonies show different responses to the ozonization process, and (ii) whether a repeated ozone exposure can enhance the inhibition of the colony growing. The results prove a high sensibility of the strains considered to the proposed treatment: the growing of both is drastically reduced, and for A. niger completely stopped. Furthermore, ozonization induces pigments bleaching in fungal colonies.     

Influence of triphenylphosphonium pendant groups on the rheological and morphological properties of new quaternized polysulfone

A new quaternized polysulfone with triphenylphosphonium pendant groups was synthesized by reacting chloromethylated polysulfone with triphenylphosphine. The molecular restructurations, generated by hydrogen bonding, electrostatic interactions, and association phenomena in ternary quaternized polysulfone/N,N‐dimethylformamide (solvent)/water (nonsolvent) systems, were evaluated by rheological investigations. The polyelectrolyte effect, induced by enhanced dissociation of the ionizable groups and by mixed solvents' quality, modify the rheological functions, that is, dynamic viscosity, elastic shear modulus, and viscous shear modulus, as well as the thermodynamic parameters obtained from the rheological properties, such as apparent activation energy. These results were correlated with the morphological properties of the films obtained from solutions in solvent/nonsolvent mixtures and compared with other quaternized polysulfones, having different hydrophobic characteristics. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013