다양한 고정화 방법에 의한 이타콘산 생산

김승욱,박승원,이진석 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.6

Aspergillus terreus NRRL 1960을 다양한 alginate gel beads, celite 및 polyurethane foam cube에 고정화하였고, 각 담체의 종류 및 크기에 따른 이타콘산의 생산을 비교하였다. Ca-alginate와 Sr-alginate에 의해 생산된 이타콘산의 농도는 비슷하였고, bentonite를 첨가했을 때(CAB bead) 생산량이 증가함을 알 수 있었다. 다양한 가교제를 alginate에 첨가했을 때 1.67% bentonite와 0.33% starch가 첨가된 Sr-alginate(SABS bead)의 경우가 다른 경우에 비해 높은 농도의 이타콘산(11.59 g/ℓ)을 생산하였다. Celite의 크기가 감소할수록 이타콘산의 생산이 증가하였으며 가장 작은 담체인 celite R-634에 균체를 고정화한 경우에 6.37 g/ℓ를 얻었다. 다양한 크기와 pore를 갖는 polyurethane foam cube를 사용했을 때, HR 08(2×2×2 ㎝)로부터 약 19 g/ℓ의 이타콘산을 얻을 수 있었고, 다른 종류의 담체에 비해 효율적이었다. Polyurethane foam cube에 고정화된 균체에 의한 반복 회분식 배양에서 4 batch까지 이타콘산을 안정하게 생산하는 것을 알 수 있었다. 또한 충진탑 반응기에서 연속배양의 가능성도 보여 주었다. Aspergillus terreus NRRL 1960 was immobilized on various alginate gel beads, Celites, and polyurethane foam cubes, and the comparisons were made for the production of itaconic acid according to the types and sizes of each carrier. The levels of itaconic acid produced from Ca-alginate and Sr-alginate were similar, and the addition of bentonite to Ca- and Sr-alginate resulted in an increase of itaconic acid. The addition of 1.67% bentonite and 0.33% starch to Sr-alginate (SABS bead) produced higher level of itaconic acid (11.59 g/ℓ) than other gel beads. A decrease in the size of Celite increased the itaconic acid production, and the smallest size of Celites, R-634, produced 6.37 g/ℓ of itaconic acid. Among various types of polyurethane foam cubes, HR08 (2×2×2 ㎝) produced about 19 g/ℓ of itaconic acid, which was more efficient than other carriers. In a repeated batch culture using immobilized cells on polyurethane foam cubes (HR08, 2×2×2 ㎝), the stability of itaconic acid production was maintained up to 4 batches. Also, the possibility of itaconic acid production by continuous culture was shown in a packed-bed reactor.

게 껍질 폐기물로부터 추훌한 Chitosan에 Itaconic acid의 그라프트 공중합과 오염물질의 제거 능력

한상문(Sang-Mun Han),윤충수(Choong-Soo Yun),김용범(Yong-Beom Kim) 유기성자원학회 1999 유기물자원화 Vol.7 No.2

수산물 가공 공장에서 배출되논 게 껍질 폐기물로부터 Chitin을 추출하고 탈아세틸화하여 Chitosan(Cs)을 제조하였다. 이 Cs를 초산 용액중에서 용해한 후 Itaconic acid를 첨가하고 CeIic ammonium nitrate(CAN)를 반응 개시제로 사용하여 Graft-copolymelization 하였다 본 연구에서는 Cs에 첨 7념F는 Itaconic acid와 CAN의 농도, 반응온도, 반응시간의 영향에 따른 최적 중합조건을 도출하고 이 Grafted-copolymel의 폐수중의 CODcr 유발물과 구리 이온의 제거 능력을 연구하였다. 여기서 CODcr과 구리이온의 제거능력 시험에 사용힌 폐수는 도금 공장 폐수를 사용하였고. Graft-copolymerization으로 합성된 이 중합체의 동정에는 적외선 스펙트로메타플 사용하였다. 이상의 실험 결과. Graft 율은 Cs에 Monomer로써 Itaconic acid 릅 0.25M농도로 첨가하였윤 때. CAN은 3.5x 10-\1. 반응 온도는 40.C 그리고 반응시간은 4시간일때 가장 높았다. 또한. Cs과 Itaconic acid grafted chitosan(Csla). Cationic polymer를 사용하여 폐수중 유기불과 금속이온의 제거 실험을 한 결과 상등액의 CODcr과 구리 이온 제거율은 Cs와 Cationic polymer 보다 CsIa가 보다 효과적이었다. 이것은 합성한 Csla가 양쪽성 고분자이기 때문이다. Chîtosan was prepared from chitin which was abstracted from wasted crab shells. Then itaconic acid was graft-copolymerized onto chitosan using ceric ammonium nitrate as a reaction initiator. To investigate the optimal grafting conditions. the influences of several factors on the grafting were studied. i.e .. the concentrations of CAN and itaconic acid. the reaction temperature and time. And to find out its flocculation ability. the flocculation test was carried out with a metal plating factory waste water. The state of graft-copolymer was identified through IR spectra analysis. The optimal grafting conditions and flocculation results were shown to be concentration of ceric ammonium nitrate is 3.5x lQ-1vi. reaction temperature is 40'c and reaction time is 4hrs with O.25M of the monomer(itaconic acid). Though flocculation tests using chitosan. grafted chitosan and cation. CODcr. metal ions removal rates were measured. The order of superiority is Itaconic acid grafted chitosan) Chitosan)Cationic polymer.

Whole-cell Immobilization of Engineered Escherichia coli JY001 with Barium-alginate for Itaconic Acid Production

문유미,Ranjit Gurav,김준영,홍윤기,Shashi Kant Bhatia,장혜림,홍주원,최태림,양수연,박형연,주황수,양영헌 한국생물공학회 2018 Biotechnology and Bioprocess Engineering Vol.23 No.4

Itaconic acid is an important organic acid and a major component of various polymers. It is used in resins, superabsorbent polymers, and substitutes for petrochemicalbased monomers such as acrylic and methacrylic acids. Itaconic acid is primarily produced by the fungus Aspergillus terreus, which yields a high titer with albeit long fermentation period and by-products. In our previous study, Escherichia coli JY001 was reported to produce itaconic acid using citric acid in whole-cell reaction resulting in higher itaconic acid productivity with less by-products formation. The present study aimed to increase whole-cell enzyme stability and reusability, via immobilization of E. coli JY001 using barium-alginate beads. We optimized the cations, temperature, pH, alginate, BaCl2 concentration, cell density per bead, and CTAB content to improve transfer rate of substrates and products. Under the optimized conditions, immobilized whole cells were stable for four repeated cycles of itaconic acid production. The present results would strengthen the basis for a continuous itaconic acid production.

Aspergillus terreus 에 의해 생합성되는 이타콘산의 생산성 증가를 위한 통계적 생산배지 최적화

장용만(Yong-Man Jang),신우식(Woo-Shik Shin),이도훈(Do-Hoon Lee),김상용(Sang-Yong Kim),박철환(Chulhwan Park),정용섭(Yong-Seob Jeong),전계택(Gie-Taek Chun) 한국생물공학회 2009 KSBB Journal Vol.24 No.1

Aspergillus terreus에 의한 이타콘산 생산 발효공정에서 생산균주의 성장을 어느 정도 제한시킴으로써 배양생리적 특성이 이타콘산 생합성 쪽으로 치우치도록 통계적 방법을 적용하여 itaconic acid의 생산배지 조성을 최적화하는 연구를 수행하였다. 이타콘산은 TCA회로를 거쳐 합성된 cis-aconitic acid의 디카르복실화 반응에 의해 생합성되는 고부가 화학원료물질이다. 우선 One factor at a time (OFAT) 방법을 이용하여 이타콘산의 생산성 증가에 크게 영향을 미치는 중요한 탄소원들로 sucrose, glucose, fructose와 soluble starch를 확인할 수 있었고, 질소원들로는 cottonseed flour와 soybean meal을 찾을 수 있었다. Fractional factorial design을 통하여 이들 6가지 요인들 간의 상호작용의 정도를 확인한 결과 sucrose와 cottonseed flour간의 상호작용의 정도가 가장 컸고, 나머지 요인들 간의 상호작용의 정도는 작거나 혹은 이타콘산 생산에 오히려 부정적인 결과를 나타냈다. 또한 full factorial design (FFD) 실험을 통해 생산배지에 KH2PO4와 MgSO4 가 과량 첨가되면 이타콘산의 생산성이 심각하게 저해됨을 알 수 있었다. FFD의 1차모델식을 근간으로 하여 최급 상승법 (steepest ascent method, SAM)을 적용하여 sucrose, cottonseed flour, KH2PO4 및 MgSO4의 최적 농도로 향하는 가장 가파른 기울기를 구함으로써, 신속하고 효율적으로 최적 농도지점에 대한 정보를 얻을 수 있었다. SAM이 제시해주는 농도 부근에서 반응표면분석 (response surface method, RSM)을 적용하여 각 배지성분의 농도를 최적화 시키기 위해, 2개의 중요한 요인인 sucrose와 cottonseed flour를 이용하여 중심합성계획 (central composite design, CCD) 실험을 수행하였다. 그 결과 이타콘산의 최적 배지조건은 sucrose 90.4 g/L, cottonseed flour 53.8 g/L인 것으로 관찰되었고, 이 농도에서 이타콘산의 생산성은 초기 사용된 배지에서의 생산성에 비해 약 7배 증가한 4360 mg/l로 나타났다. 이로부터 탄소원 (C)으로 사용한 sucrose와 질소원 (N)으로 사용한 cottonseed flour 간의 C/N 비율이 이타콘산의 생산성에 큰 영향을 미친다는 것을 확인할 수 있었다. Statistical optimization of the production medium was carried out in order to find an optimal medium composition in itaconic acid fermentation process. Itaconic acid utilized in the manufacture of various synthetic resins is a dicarboxylic acid biosynthesized by fungal cells of Aspergillus terreus in a branch of the TCA cycle via decarboxylation of cis-aconitate. Through OFAT (one factor at a time) experiments, six components (glucose, fructose, sucrose, soluble starch, soybean meal and cottonseed flour) were found to have significant effects on itaconic production among various carbon- and nitrogen-sources. Hence, using these six factors, interactive effects were investigated via fractional factorial design, showing that the initial concentrations of sucrose and cottonseed flour should be high for enhanced production of itaconic acid. Furthermore, through full factorial design (FFD) experiments, negative effects of KH2PO4 and MgSO4 on itaconic acid biosynthesis were demonstrated, when excess amounts of the each component were initially added. Based on the FFD analysis, further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to obtain optimal concentrations of the constituent nutrients. As a result, optimized concentrations of sucrose and cottonseed flour were found to be 90.4g/L and 53.8g/L respectively, with the corresponding production level of itaconic acid to be 4.36 g/L (about 7 fold higher productivity as compared to the previous production medium). From these experimental results, it was assumed that optimum ratio of the constituent carbon (sucrose) and nitrogen (cottonseed flour) sources was one of the most important factors for the enhanced production of itaconic acid.

Surface conjugation of poly (dimethyl siloxane) with itaconic acid-based materials for antibacterial effects

Birajdar, Mallinath S.,Cho, Hyunjoo,Seo, Youngmin,Choi, Jonghoon,Park, Hansoo Elsevier BV * North-Holland 2018 Applied Surface Science Vol.437 No.-

<P><B>Abstract</B></P> <P>Poly (dimethyl siloxane) (PDMS) is widely used in various biomedical applications. However, the PDMS surface is known to cause bacterial adhesion and protein absorption issues due to its high hydrophobicity. Therefore, the development of antibacterial and anti-protein products is necessary to prevent these problems. In this study, to improve its antibacterial property and prevent protein adsorption, PDMS surfaces were conjugated with itaconic acid (IA) and poly (itaconic acid) (PIA) via a chemical method. Additionally, IA and PIA were physically blended with PDMS to compare the antibacterial properties of these materials with those of the chemically conjugated PDMS surfaces. The successful synthesis of the PIA polymer structure was confirmed by proton nuclear magnetic resonance (<SUP>1</SUP>H NMR) spectroscopy. The successful conjugation of IA and PIA on PDMS was confirmed by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), water contact angle measurements, and microbicinchoninic acid (BCA) protein assay analyses. The PDMS surfaces functionalized with IA and PIA by the conjugation method better prevented protein adsorption than the bare PDMS. Therefore, these surface-conjugated PDMS can be used in various biomedical applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A best bacterial anti-adhesion system was developed based on itaconic acid hydrophilic materials via chemical conjugation and physical blending methods. </LI> <LI> Itaconic acid (IA) and poly (itaconic acid) (PIA) were conjugated chemically onto PDMS surfaces to improve antibacterial activity. </LI> <LI> The PDMS surfaces functionalized with IA and PIA increased contact angle, prevented protein adsorption than the bare PDMS and also did not show cytotoxicity. </LI> <LI> The best bacterial anti-adhesion effect was observed for the high-concentration PIA-conjugated PDMS surface prepared by the combined method. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Aspergillus itaconicus 유래 itaconic acid의 ester-monomer를 이용한 새로운 soft contact lens 소재 개발

유영현(Young-Hyun You),남주형(Joo-Hyeung Nam),김병길(Bieong-Kil Kim),김순복(Soon bok Kim),문익재(Ik-Jae Moon),김종필(Jong-Pil Kim),서영배(Young-Bae Seu) 한국생명과학회 2009 생명과학회지 Vol.19 No.4

본 연구는 itaconic acid로부터 합성한 새로운 soft contact lens 소재의 함수율과 산소투과도를 확인 하였다. Soft contact lens의 새로운 polymer소재 개발을 위하여 기본 monomer인 HEMA (2-hydroxyethyl methacrylate)와 가교제로서 EGDMA (ethylene glycol dimethacrylate) 그리고 A. itaconicus의 발효에 의해 생산된 itaconic acid로부터 얻어진 mono-ester 1과 di-ester 2를 새로운 첨가제로 사용하여 중합반응을 시도하였다. Soft contact lens의 새로운 폴리머 소재들은 HEMA와 mono-ester 1 또는 di-ester 2를 서로 다른 비율로 합성하여 함수율(water content)과 산소투과도(oxygen permeability)를 분석하였다. Polymer 형성을 위한 HEMA와 additive의 혼합에서 HEMA/mono-ester 1의 혼합물은 HEMA/di-ester 2 보다 함수율과 산소투과도가 뛰어 났으며 HEMA와 첨가제 mono-ester 1 (15%)를 이용한 중합의 경우, 제작된 soft contact lens의 함수율과 산소투과도는 각각 57.7%와 28.6 Dk로 좋은 값을 나타냈다. 새로운 소재를 사용하여 만든 soft contact lens의 함수율과 산소투과도는 우수하게 나타났다. In this study, we confirmed water content and oxygen permeability of new polymeric materials synthesized from itaconic acid used for soft contact lenses. In this study, we polymerized materials for soft contact lenses using HEMA (2-hydroxyethyl methacrylate), the based-monomer of soft contact lenses, EGDMA (ethylene glycol dimethacrylate) as a cross linkage agent, and the new additives mono-ester or di-ester derived from itaconic acid commercially produced by the fermentation of A. itaconicus. New polymer materials for contact lenses were synthesized with the mixture of HEMA and mono- or di-ester at different ratios and water content and oxygen permeability (Dk) was analyzed. In polymerizing HEMA and mono-ester (15%), the water content and oxygen permeability of contact lenses were found to be of good value at 57.7% and 28.6 Dk respectively. The mixture of HEMA and mono-ester is more excellent than HEMA/di-ester in regards to water content and oxygen permeability. The water content and oxygen permeability of soft contact lenses made by new polymeric materials were highly represented.

Poly(acrylic acid-co-itaconic acid) for dental glass ionomer cement with improved properties

조동원,장홍규,홍성철 한국공업화학회 2016 한국공업화학회 연구논문 초록집 Vol.2016 No.1

Glass ionomer cement (GIC) has been widely used in the dental field because of its fluoride release, chemical adhesion to tooth structure, and available to use in a variety of clinical scenarios. However, the major disadvantages of current glass ionomer cement are short storage period and long curing time. In this study, a well-defined poly(acrylic acid-co-itaconic acid) (PAI) is prepared with controlled composition and molecular weight. The resulting PAI copolymers are investigated by using proton nuclear magnetic resonance spectroscopy, infrared spectroscopy and gel permeation chromatography. The PAI copolymers are further used as an acid part in GIC. The outstanding storage stability is achieved by using the PAI copolymers. The curing time is also controlled by the formulation of GIC.

Itaconic Acid-based Superabsorbent Polymer Composites Using Cellulose with Enhanced Absorption Properties and Heat Resistance

Hae Chan Kim,Seung Ho Lim,Yong Rok Kwon,김정수,Jong-Ho Kim,Dong Hyun Kim 한국섬유공학회 2022 Fibers and polymers Vol.23 No.4

The polymerization of itaconic acid is generally characterized by a slow polymerization rate and poor networkformation. This results in a polymer with low absorption properties. To improve absorption properties and for use in thehygiene industry, we have been researching superabsorbent polymers (SAPs) based on itaconic acid precursors. In this study,poly(itaconic acid-co-acrylic acid) composites using cellulose, a natural polymer, were synthesized by aqueous solutionpolymerization, and the surface-crosslinking of the SAP composites was introduced. We characterized the structure of SAPcomposites using Fourier transform infrared (FT-IR) spectroscopy. We also measured absorption properties such ascentrifuge retention capacity (CRC), absorbency under load (AUL), and permeability in 0.9 wt.% saline solution. Weconfirmed the synthesis conditions of SAP composites by crosslinker type, cellulose content, and surface-crosslinkingconditions. We found the best performance of an SAP with a CRC of 27.8 g/g, AUL of 21.1 g/g, and permeability of 61 s at5 g input cellulose. In addition, surface characteristics were evaluated by scanning electron microscopy, and thermalstabilities were analyzed by thermo gravimetric analysis.

Enhancing conversion of acetate into itaconic acid with engineered Escherichia coli

김민재,노명현,우성화,임현규,정규열 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-

Utilization of cheap carbon sources are both beneficial to production cost and economic feasibility. In the previous study, production of itaconate from acetate was constructed with rational metabolic engineering strategy, proving the potential of acetate as a promising carbon source. In this study, we introduced combinatorial approach to further increase the production of itaconate. Initially, the itaconate-specific screening device could be constructed using transcriptional regulator and selection marker. Thus, Adaptive Laboratory Evolution (ALE), utilizing natural selection toward microorganisms under selection pressure, was introduced to itaconate-producing strain. The evolved strain could produce 1.06 g/L of itaconate, a 1.24-fold increase in comparison with parental strain and the yield was significantly enhanced by 7.16-fold. The rational approach will be applied to maximize the itaconate production and the mutant will be additionally anaylzed with WGS for validation.

A novel approach for itaconic acid extraction: Mixture of trioctylamine and tridodecylamine in different diluents

Yavuz Selim Asci,Ismail Inci 한국공업화학회 2012 Journal of Industrial and Engineering Chemistry Vol.18 No.5

Extraction of itaconic acid has been investigated from aqueous solutions by six kind of different solutions of trioctylamine (TOA)–tridodecylamine (TDA) mixtures. Solvents have selected as dimethyl phthalate (DMP), methyl isobutyl ketone (MIBK), 2-octanone, 1-octanol, cyclohexyleacetate (CHA), and 1-decanol. Distribution and equilibrium data for design of separation equipments have been obtained as a result of batch extraction of itaconic acid. It has been calculated that distribution coefficients (D), loading factors (Z), extraction efficiency (E). The maximum removal of itaconic acid has been obtained as 98.39% with DMP and 3.14 mol L-1 initial concentration of TOA–TDA mixture.