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Bacterial Cellulose Production by Acetobacter Xylinum ATCC 23767 Using Tobacco Waste Extract As Culture Medium T

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Bacterial Cellulose Production by Acetobacter Xylinum ATCC 23767 Using Tobacco Waste Extract As Culture Medium T Bioresource Technology 274 (2019) 518–524 Contents lists available at ScienceDirect Bioresource Technology journal homepage: www.elsevier.com/locate/biortech Bacterial cellulose production by Acetobacter xylinum ATCC 23767 using tobacco waste extract as culture medium T Jianbin Yea, Shanshan Zhenga, Zhan Zhangb, Feng Yangc,KeMaa, Yinjie Fengb, Jianqiang Zhenga, ⁎ Duobin Maoa, Xuepeng Yanga, a School of Food and Biological Engineering, Henan Provincial Collaborative Innovation Center for Food Production and Safety, Zhengzhou University of Light Industry, Dongfeng Road 5#, Zhengzhou 450002, Henan Province, China b Technology Center, China Tobacco Henan Industrial Co., Ltd, Zhengzhou 450000, China c Henan Cigarette Industrial Tobacco Sheet Co, Ltd, Henan, Xuchang 461000, China GRAPHICAL ABSTRACT ARTICLE INFO ABSTRACT Keywords: In this study, bacterial cellulose (BC) was synthesized by Acetobacter xylinum ATCC 23767 using tobacco waste Bacterial cellulose extract (TWE) as a carbon source. Nicotine was found to be an inhibitory factor for BC synthesis, but it can be Tobacco waste extract removed at pH 9.0 by steam distillation. After removing nicotine, the BC production was 2.27 g/L in TWE Acetobacter xylinum prepared with solid-liquid (S-L) ratio at 1:10. To further enhance the BC production, two fermentation stages Nicotine were performed over 16 days by re-adjusting the pH to 6.5 at 7 days, after the first fermentation stage was Fermentation completed. Using this two-stage fermentation, the BC production could reach 5.2 g/L. Structural and thermal analysis by FE-SEM, FT-IR, XRD and TGA showed the properties of BC obtained from TWE were similar to that from Hestrin-Schramm (HS) medium. Considering the huge disposal tobacco waste in China, the present study provides an alternative methodology to synthesize BC. 1. Introduction glycosidic bonds. Traditionally, cellulose is extracted from the lig- nocellulosic biomass of plants, but it can also be synthesized by some Cellulose is one of the most abundant natural polymers on earth and bacteria, such as Gluconacetobacter, Acetobacter, Agrobacterium, and has been widely used in many fields (Cacicedo et al., 2016; Jahan et al., Rhizobium (Uzyol and Sacan, 2017). The latter material is called bac- 2018). It is a polymer of glucose units linked together by β-1, 4- terial cellulose (BC). Although it has similar chemical composition to ⁎ Corresponding author. E-mail address: [email protected] (X. Yang). https://doi.org/10.1016/j.biortech.2018.12.028 Received 5 November 2018; Accepted 9 December 2018 Available online 10 December 2018 0960-8524/ © 2018 Elsevier Ltd. All rights reserved. J. Ye et al. Bioresource Technology 274 (2019) 518–524 plant cellulose, BC has higher degrees of purity, tensile strength, crys- liquid pretreatment in the previous studies (Cheng et al., 2017; Hong tallinity, porosity, polymerization, water-holding capacity, biodegrad- et al., 2012). However, only the free sugar was extracted and utilized ability, and biological adaptability (Campano et al., 2016; Wang et al., for the BC production in the present study, as the rest part of solid was 2017). These superior physicochemical properties result in a wide range transferred to the Reconstituted Tobacco Company of Xuchan (Henan of application, such as in the biomedical, textile, audio device (Lima Province, China) for the production of reconstituted tobacco. Acet- et al., 2017; Shah et al., 2013; Shi et al., 2014). Thus, BC synthesis has obacter xylinum ATCC 23767 was purchased from the Institute of Mi- attracted the increasing interest of many researchers. crobiology, Chinese Academy of Sciences. Nicotine was purchased from However, the high production cost of BC has become one of the Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). Plant cel- main drawbacks to its general application in industrial and academic lulose standard (CAS, 9004-34-6) and other chemicals were obtained fields (Hu et al., 2010). The typical substrate for BC production includes from Shanghai Aladdin Bio-chem Technology Co., Ltd (Shanghai, several types of carbon sources, such as glucose, sucrose, fructose, China). glycerol, mannitol, and arabitol (Hong et al., 2012; Mohammadkazemi et al., 2015). The traditional Hestrin–Schramm (HS) medium was 2.2. Methods commonly used for the BC production, which is expensive and requires additional products, such other carbon source, yeast or peptone, etc. 2.2.1. Preparation of TWE and other culture media Therefore, inexpensive raw materials containing high levels of reducing TWE was prepared as follows. (1) Tobacco waste was boiled with sugars are frequently considered promising substrates for BC produc- distilled water for 1.5 h at different solid-to-liquid ratios (SL ratios, w/v, tion. Recently, various industrial by-products and agroforestry wastes 1:4, 1:6, 1:8, 1:10, or 1:12). (2) After cooling, insoluble solids were have been utilized as carbon sources to improve the yield and reduce removed and the TWE was collected by filtering through muslin cloth the cost of BC production. Thus far, several raw waste materials have under vacuum. (3) The pH of the TWE was adjusted to 6.5 by adding been demonstrated to be potential substrates for BC production, such as 0.2 M NaOH or HCl. The sugar yield (%) was defined as Eq. (1), and the distillery effluent (Jahan et al., 2018), corn steep liquor (Costa et al., volume (L) was according to the liquid used during the extraction. 2017), fruit juice (Kim et al., 2017), corn stalks (Cheng et al., 2017), litchi extract (Yang et al., 2016), beverage industrial waste (Fan et al., Sugar yield (%) 2016), corncob acid hydrolysate (Huang et al., 2015), and waste beer Total sugar concentration in TWEmg ∗∗ volume( L ) 1000 ml ()L ()L yeast (Lin et al., 2014). = solid tobacco waste weight Tobacco (Nicotiana) is an economic crop cultivated worldwide (mg) (Wang et al., 2015). China is the largest consumer and producer of ∗ 100% (1) fl tobacco products in the world, with more than 2 million tons of ue- To investigate the effect of extra carbon sources on BC production, cured tobacco used for cigarette production every year (Liu et al., modified TWE media were prepared by adding 5 g/L glucose, fructose, 2015). During the production process, approximately 1 million tons of sucrose, or mannose. To evaluate the effect of nicotine on BC produc- tobacco waste is produced, which includes unwanted tobacco leaves, tion, Hestrin-Schramm (HS) medium was prepared as follows (g/L): tobacco stems, and scraps (Wang et al., 2013; Zhong et al., 2010). glucose (20.0), peptone (5.0), yeast extract (5.0), Na HPO (2.7), and fi 2 4 Unfortunately, this tobacco waste is commonly dumped into land lls or citric acid monohydrate (1.15), pH adjusted to 6.5. Different con- incinerated because of its high content of toxic nicotine (Okunola et al., centrations of nicotine (0, 0.4, 0.6, 0.8, 1.2, 1.6, 2.0, or 2.4 g/L) were 2016; Zhang et al., 2013). This has endangered human health and added to the HS medium. Pre-culture medium was prepared to increase contributed to environmental pollution. Recently, tobacco waste has the population of bacteria before fermentation (g/L): glucose (20.0), been used as a substrate to produce fertilizer, pectinase, and some peptone (5.0), and yeast extract (5.0), initial pH of 6.5. medicine precursors, for example (Chaturvedi et al., 2008; Wang et al., 2015; Zheng et al., 2016, 2017). However, to our knowledge, there are 2.2.2. Remove of nicotine by steam distillation no reports showing tobacco waste as a substrate for BC production. In The nicotine in TWE was removed by steam distillation at different fact, the high content of sugars present in tobacco waste, including pH values. After ensuring the optimal solid-to-liquid ratio, 1000 mL of glucose, sucrose, fructose, and other polysaccharides, would make to- prepared TWE was added to the distillation flask, and the pH adjusted bacco waste a promising substrate for BC production. to various values (3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, or 10.0) by addition of In this study, we first evaluated tobacco waste extract (TWE) as a NaOH or HCl. Then, 200 mL of liquid was removed by steam distillation substrate for BC production and demonstrated that nicotine in the to eliminate the nicotine from the TWE. After cooling, 200 mL deio- medium is an inhibitor of this process. Then, a steam-distillation pro- nized water was added to the flask to replenish the volume and the pH cess was used to remove the nicotine and increase the BC yield. BC was returned to 6.5 by the addition of NaOH or HCl. The original and production was further increased by adjusting the pH after first-stage residual nicotine contents of the TWE were both detected by high fermentation. Using this two-stage fermentation strategy, TWE was performance liquid chromatography (HPLC), and the nicotine removal shown to be an ideal substrate for BC production, which has the po- was calculated as equation (2): tential to reduce the cost of BC production and to solve the problem of tobacco waste. This study provides a green and sustainable method to Nicotine removal rate reuse tobacco waste. original nicotine content- residual nicotine content =∗100% original nicotine content (2) 2. Materials and methods 2.1. Materials 2.2.3. Strain incubation and BC production A. xylinum ATCC 23767 powder was activated in a 10-mL tube Tobacco waste, was collected from China Tobacco Henan Industrial containing pre-culture medium as described above at 30 °C for 2 days Co., Ltd. (Zhengzhou, China). The waste was found to be composed of with 150 rpm shaking. The activated bacteria were then cultured for 25.4% cellulose, 22.3% hemicellulose, 3.1% lignin, 6.2% pectin, and 2 days in a 250-mL shaker bottle containing 100 mL HS medium to ∼38.5% soluble substance and 5.1% ash.
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