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Artículo Ácido Lactico
Artículo Ácido Lactico
Artículo Ácido Lactico
Process Biochemistry
journal homepage: www.elsevier.com/locate/procbio
Keywords: Sugarcane molasses, a waste from sugar manufacturing processes, has promising future to be utilized as a cheap
Microbial carbon source for lactic acid production. In this study, a newly microbial consortium, CEE-DL15, for the con-
Consortium version of sugarcane molasses to lactic acid was selected and evaluated. The consortium was screened from cattle
Lactic acid stomach content and mainly consisted of Clostridium sensustricto (57.29%), Escherichia (34.22%), and
Sugarcane molasses
Enterococcus (5.32%). Lactic acid production was explored under the deficiency of sterilization and molasses
Corn steep liquor powder
acidification, with corn steep liquor powder used as organic nitrogen source. In batch fermentations using su-
garcane molasses of 350 g/L and corn steep liquor powder of 18.5 g/L without additional nutrients, CEE-DL15
produced 112.34 g/L lactic acid (107.40 g/L L-lactic acid and 4.94 g/L D-lactic acid), with a yield of 0.81 g/g and
a maximum productivity of 4.49 g/(L.h), which is the best lactic acid productivity from molasses published so
far. Economic analysis indicated that lactic acid fermentation cost was only 448 USD/ton using molasses and
corn steep liquor powder, which was only 37.7% of the total cost when compared with glucose as carbon source
and MRS medium as nitrogen source. This work demonstrated that the high adaptation to molasses of microbial
consortium CEE-DL15 might be a promising alternative for the economical production of lactic acid.
1. Introduction available raw material for microbial production [10]. Sugarcane mo-
lasses contains about 50% (w/v) total sugar (sucrose, glucose, and
Lactic acid is an important platform chemical that has been applied fructose), 0.5−0.9% (w/v) nitrogen, 10% (w/v) inorganic salts, etc
in foods, cosmetics, textiles, pharmaceuticals and many other industrial [11,12]. However, some hazardous substances such as 5-hydro-
fields. Biotechnological lactic acid production has several advantages xymethylfurfural and excessive metallic ions are generated during
over chemical approaches regarding environmental issues, good utili- sugar manufacturing process, which are toxic to cell growth, causing
zation of diverse substrates, and the feasibility to produce optically pure low conversion yield and productivity [13]. To eliminate the inhibition
lactic acid forms [1]. However, higher production costs have hindered of hazardous substances and improve sugarcane molasses utilization,
the large-scale application of lactic acid. Therefore, reduction of lactic genetic engineering strains or mutagenic strains were applied to lactic
acid production cost through utilization of inexpensive substrate and acid production [14,15]. Escherichia coli WYZ-L, enhancing expression
improvement of lactic acid production and productivity has become an of the sucrose operon (cscA and cscKB), produced 75 g/L of lactic acid,
important goal [2]. with a yield of 0.85%, and a maximum productivity of 1.18 g/(L.h)
The carbon source in lactic acid fermentation is responsible for the using sugarcane molasses and corn steep liquor without additional
major factors in the economic production of lactic acid. Studies on nutrients [15]. High lactic acid concentration of 166 g/L was obtained
lactic acid fermentation from raw materials as fermentative substrates at a molasses sugar concentration of 190 g/L with a productivity of
have suggested numerous candidates, such as sugarcane molasses [3,4], 4.15 g/(L.h) by Lactobacillus delbrueckii mutant Uc-3, isolating by UV
paper sludge [5], cellulose [6], apple pomace [7], kitchen wastes [8], mutagenesis [14]. Moreover, co-feeding strategy based on the utiliza-
Jerusalem artichoke powder [9], etc. As a relatively cheap and abun- tion of cane molasses/glucose carbon sources was considered to im-
dant raw material, sugarcane molasses is a by-product prepared from prove the utilization of cane molasses. A titer of 168.3 g/L L-lactic acid
the liquid waste of sugar production and the output of sugarcane mo- was obtained by a Bacillus coagulans strain H-1 after 78 h fed-batch
lasses in China is about 3 million tons annually, which makes it an fermentation, with a productivity of 2.1 g/(L.h) and a yield of 0.88 g/g
⁎
Corresponding author at: School of Life Science and Biotechnology, Dalian University of Technology, No. 2 Linggong Road, Ganjingzi District, Dalian City,
Liaoning Province, 116024, PR China.
E-mail address: zhlxiu@dlut.edu.cn (Z. Xiu).
https://doi.org/10.1016/j.procbio.2019.03.022
Received 31 October 2018; Received in revised form 7 February 2019; Accepted 24 March 2019
1359-5113/ © 2019 Elsevier Ltd. All rights reserved.
Please cite this article as: Yaqin Sun, et al., Process Biochemistry, https://doi.org/10.1016/j.procbio.2019.03.022
Y. Sun, et al. Process Biochemistry xxx (xxxx) xxx–xxx
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Table 3
The concentration of lactic acid isomers at different sugarcane molasses con-
centrations.
Samples Sugarcane Fermentation Total L-lactic D-lactic L-lactic
molasses(g/L) time (h) lactic acid acid acid
acid (g/L) (g/L) (g/L) (%)
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Table 4
Costs analysis and comparison for carbon source, nitrogen source and sterilization.* *Costs and energy consumption were per ton lactic acid produced.
References Lactic acid Carbon source Nitrogen source Sterilization Material and
(US dollar $) (US dollar $) sterilization
Concentration Yield Productivity Energy Cost (US dollar $)
(g/L) (g/g) (g/(L.h)) consumption (US dollar $)
(106 kJ)
[29] 192.0 0.96 3.99 Glucose 521 yeast extract (5 g/L), peptone 619 1.97 49 1189
(10 g/L),
beef extract (10 g/L)
[14] 166.0 0.95 4.20 Molasses 263 yeast extract (5 g/L) 179 2.28 57 499
[4] 165.7 0.92 1.60 Glucose 546 CSLP (1 g/L) 3 2.28 57 606
168.3 0.88 2.10 Glucose/ 446 CSLP (1 g/L) 3 2.25 56 505
Molasses
[15] 75.0 0.85 1.18 Molasses 294 CSLP -a 5.04 125 419b
[28] 107.0 0.90 1.48 Molasses 278 yeast extract (5 g/L), peptone 619 3.53 88 985
(10 g/L),
beef extract (10 g/L)
This study 112.3 0.81 4.49 Molasses 375 CSLP (18.5 g/L) 73 0 0 448
Costs and energy consumption were per ton lactic acid produced.
a
The mass of CSLP was not given.
b
The cost of nitrogen source was not included.
an enhanced expression of the sucrose operon (cscA and cscKB) [15]. Table 4 showed that the cost of cane molasses occupied almost 50–60%
High concentration of lactic acid (166 g/L) was obtained with a pro- portion of the cost of glucose in different studies [14,28,29]. The cost
ductivity of 4.2 g/(L.h) by Lactobacillus delbrueckii mutant Uc-3 [14]. for MRS medium including yeast extract, peptone and beef extract oc-
The mutant was isolated by UV mutagenesis followed by selection on cupied over 50% portion of total cost for materials and sterilization
the basis of a bigger zone of acid formation on sucrose-based medium. [28,29], while the cost of CSLP was only 10% compared to using MRS
In their studies, sugarcane molasses was pretreated by hydrolysis and as nitrogen source. Economic analysis indicated that lactic acid pro-
5 g/L yeast extract was used to supply nitrogen. It is noteworthy that duction cost was 1189 USD/ton using glucose as carbon source and
only CSLP was required to obtain high lactic acid productivity even at a MRS medium as nitrogen source. Lactic acid production cost was 499
high molasses concentration according to our work. A maximum pro- USD/ton using molasses as carbon source and yeast extract as nitrogen
ductivity of 4.49 g/(L.h) was obtained, which is the best lactic acid source by Lactobacillus delbrueckii subsp. delbrueckii mutant Uc-3 in
productivity from molasses published so far. batch fermentation. In our study, lactic acid production cost was only
According to previous studies, the primary cost of lactic acid pro- 448 USD/ton using molasses as carbon source and CSLP as nitrogen
duction includes fermentative substrates of nutrients, nitrogen sources source by microbial consortium CEE-DL15, which is only 37.7% of the
and carbon sources [2]. In this study, the cost of carbon sources and cost when using glucose as the solo carbon source and MRS medium as
nitrogen sources was analyzed to evaluate the batch fermentation for nitrogen source. Therefore, lactic acid production strategy from su-
lactic acid production by microbial consortia CEE-DL15. The best re- garcane molasses and CSLP by microbial consortium CEE-DL15 is pre-
ported results of lactic acid production from glucose and molasses were ferable on an economical viewpoint.
selected for comparison with our research [14,29] and the results were It should be pointed out that sugarcane molasses should be con-
presented in Table 4. trolled by acidification according to the study of Dumbrepatil A et al
The cost of carbon and nitrogen sources was defined as [14]. Cane molasses contains a number of inorganic and organic in-
hibitors, which may impede the metabolism of cells. Therefore, cane
Ci Pi
CLA
( i = cs, ns ) , where Ccs , Cns and CLA represent the concentrations of
carbon source, nitrogen sources in medium and lactic acid produced, molasses needs to be pretreated to reduce the level of various inhibitors.
respectively. Pcs and Pns represent the prices of carbon source and ni- However, pretreatment will raise the fermentation cost of cane mo-
trogen source. Costs and energy consumption were per ton lactic acid lasses and increase workload. In the present study, sugarcane molasses
produced. The prices of glucose and cane molasses were approximately was used without any pretreatment, which might be due to the strong
500 USD/ton and 120 USD/ton, respectively. The prices of nitrogen tolerance of microbial consortium CEE-DL15. Moreover, the char-
sources in MRS medium consisting of yeast extract, peptone and beef acteristic of open fermentation ability owned by microbial consortium
extract were approximatly 5944 USD/ton, 4458 USD/ton and 4458 CEE-DL15 makes the fermentation medium be utilized without auto-
USD/ton, respectively. The price of corn steep liquor powder in our claving.
study was approximately 446 USD/ton. Moreover, the cost of ster-
ilization was also analyzed and evaluated. The energy consumption of
sterilization was defined as Q = C*M*Δt, where C, M, andΔt represent 4. Conclusion
the specific heat of fermentation broth, mass of fermentation broth and
temperature difference, respectively. For simplicity, the specific heat of In this study, we successfully achieved high production of lactic acid
fermentation broth was assigned as 4.2 kJ/(kg.oC), the specific heat of and high tolerance of molasses with a selected microbial consortium
water. Costs and energy consumption in Table 4 were per ton of re- CEE-DL15. The evaluation of CEE-DL15 has the potential for cost-ef-
quired lactic acid produced. fective production of lactic acid using CSLP as medium without addi-
When using cane molasses and glucose as co-feeding carbon sources, tional nutrients supplements. 112.34 g/L Lactic acid (107.40 g/L L-
the production cost was 83% compared to using glucose as the solo lactic acid and 4.94 g/L D-lactic acid), with a yield of 0.81 g/g, a pro-
carbon source in the study of Xu et al [4]. Although the low pro- ductivity of 4.49 g/(L.h), which is the best lactic acid productivity from
ductivity was obtained in their study, cane molasses used in this study molasses published so far was obtained when 350 g/L sugarcane mo-
was not pretreated, and the organic nitrogen in the fermentation lasses were utilized without any pretreatment and sterilization. This
medium was only 1 g/L corn steep powder. The cost analysis and study provides an efficient and economical option for the utilization of
comparison for carbon source, nitrogen source and sterilization in raw materials in microbial production.
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