去乙酰化槐糖脂生物表面活性剂的结构鉴定和理化性质

doi:10.19894/j.issn.1000-0518.240021

摘要/Abstract

摘要：

槐糖脂亲水基上的乙酰化程度对其表界面活性和理化性质有重要影响，为了查明不同菌株发酵得到的乙酰化/去乙酰化槐糖脂（SLs）在结构和理化性能上的差异，使用HPLC-MS/MS解析了去乙酰化SLs的结构，并与野生菌株所产乙酰化SLs的表界面活性等理化性质进行比较。结果表明：去乙酰化菌株所产的去乙酰化SLs疏水基主要为十八烯酸，亲水基主要为去乙酰化槐糖，其中，内酯型、酸性和具有bola型结构的槐糖脂质量分数分别为26.99%、49.98%和23.03%。野生菌株所产槐糖脂以乙酰化槐糖脂为主，且内酯型和酸型质量分数分别为97.86%和2.14%；其疏水基主要为十八烯酸。去乙酰化SLs在水中的溶解度高达485.8 g/L，较乙酰化SLs提高了14倍，且具有更好的发泡性能和泡沫稳定性，同时，去乙酰化SLs乳化性能较乙酰化SLs提高26.7倍。去乙酰化SLs的表面活性稍低，临界表面张力为41.0 mN/m，高于乙酰化SLs的36.2 mN/m；亲水-亲油平衡值为13，高于乙酰化SLs的11。 2种槐糖脂具有良好的抗硬水性，但耐酸碱性均较弱。

关键词: 假丝酵母菌, 槐糖脂, 去乙酰化, 临界胶束浓度, 乳化性

Abstract:

Sophorolipids （SLs） are a kind of biosurfactants with the highest potential for industrial application， and the degree of acetylation in their sugar chains has an important influence on their surface interface activity，and physicochemical properties. By homologous recombination， the research group constructed the acetyltransferase gene-deficient strain， and nonacetylated SLs was prepared by fermentation with oleic acid and glucose as biphasic carbon sources. In this paper， the structure of nonacetylated SLs was analyzed by high-performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）， and the physicochemical properties of acetylated SLs produced by wild strains were compared. The results showed that the hydrophobic groups of nonacetylated SLs produced by nonacetylated strains were mainly octadecenoic acid， and the hydrophilic groups were mainly nonacetylated sophoricoside， in which the contents of lipoid， acidic and bola-type SLs were 26.99%， 49.98% and 23.03% respectively. The sophorolipids produced by wild strains are mainly acetylated sophorolipids， and the mass fractions of lactone and acid types are 97.86% and 2.14% respectively. Its hydrophobic group is mainly octadecenoic acid. The solubility of nonacetylated SLs in water is as high as 485.8 g/L， which is 14 times higher than acetylated SLs， and it has better foaming performance and foam stability. At the same time， the emulsifying performance of nonacetylated SLs is 26.7 times higher than acetylated SLs. Due to the increase of hydrophilicity of the missing acetyl group， the surface activity of deacetylation SLs decreased， and the surface tension of aqueous solution was reduced to 41.0 mN/m， which was higher than that of acetylated sophorolipid （36.2 mN/m）. The hydrophilic and lipophilic balance value is 13， which is higher than that of acetylation SLs （11）. Two kinds of sophorolipid have good hard water resistance， but weak acid and alkali resistance.

Key words: Candida, Sophorolipid, Deacetylation, Critical micelle concentration, Emulsibility

中图分类号:

O647.2

杨昊, 马孝萌, 黄金华, 李俊峰, 梁生康. 去乙酰化槐糖脂生物表面活性剂的结构鉴定和理化性质[J]. 应用化学, 2024, 41(6): 878-889.

Hao YANG, Xiao-Meng MA, Jin-Hua HUANG, Jun-Feng LI, Sheng-Kang LIANG. Structural Identification and Physicochemical Properties of Nonacetylated Sophorolipids Biosurfactant[J]. Chinese Journal of Applied Chemistry, 2024, 41(6): 878-889.

图/表 11

参考文献 33

1	PAL S， CHATTERJEE N， DAS A K， et al. Sophorolipids： a comprehensive review on properties and applications‍‍［J］. Adv Colloid Interface Sci， 2023， 313： 102856.
2	MADANKAR C S， BORDE P K. Review on sophorolipids-a promising microbial bio-surfactant‍‍［J］. Tenside Surfact Det， 2023， 60（2）： 95-105.
3	CHO W Y， NG J F， YAP W H， et al. Bio-based antimicrobial formulating agents for applications in food and health‍［J］. Molecules， 2022， 27（17）： 5556.
4	DEVELTER D W G， LAURYSSEN L M L. Properties and industrial applications of sophorolipids‍‍［J］. Eur J Lipid Sci Technol， 2010， 112（6）： 628-638.
5	ASMER H J， LANG S， WAGNER F， et al. Microbial production， structure elucidation and bioconversion of sophorose lipids［J］. J Am Oil Chem Soc， 1988， 65（9）： 1460-1466.
6	DENGLE PULATE V， BHAGWAT S， PRABHUNE A. Microbial oxidation of medium chain fatty alcohol in the synthesis of sophorolipids by candida bombicola and its physicochemical characterization‍［J］. J Surfactants Deterg， 2013， 16（2）： 173-181.
7	KONISHI M， FUKUOKA T， MORITA T， et al. Production of new types of sophorolipids by by candida batistae‍［J］. J Oleo Sci， 2008， 57（6）： 359-369.
8	VAN BOGAERT， SAERENS K， DE MUYNCK C， et al. Microbial production and application of sophorolipids‍［J］. Appl Microbiol Biot， 2007， 76（1）： 23-34.
9	SEN A R， KARIM S A， SANTRA A. Effect of defaunation on carcass and meat characteristics of finisher lambs‍［J］. Indian J Anim Sci， 2000， 70（6）： 659-661.
10	OTTO R T， DANIEL H J， PEKIN G， et al. Production of sophorolipids from whey‍［J］. Appl Microbiol Biot， 1999， 52（4）： 495-501.
11	LI S， QIAN X， XU L， et al. Biological tailoring of novel sophorolipid molecules and their derivatives‍［J］. Biofuel Bioprod Bior， 2021， 15（6）： 1938-1949.
12	SHAO L， SONG X， MA X，et al. Bioactivities of sophorolipid with different structures against human esophageal cancer cells［J］. J Surg Res， 2012， 173（2）： 286-291.
13	RAU U， HECKMANN R， WRAY V， et al. Enzymatic conversion of a sophorolipid into a glucose lipid‍［J］. Biotechnol Lett， 1999， 21（11）： 973-977.
14	BORSANYIOVA M， PATIL A， MUKHERJI R， et al. Biological activity of sophorolipids and their possible use as antiviral agents［J］. Folia Microbiol， 2016， 61（1）： 85-89.
15	KASTURE M， SINGH S， PATEL P， et al. Multiutility sophorolipids as nanoparticle capping agents： synthesis of stable and water dispersible Co nanoparticles［J］. Langmuir， 2007， 23（23）： 11409-11412.
16	LYDON H L， BACCILE N， CALLAGHAN B， et al. Adjuvant antibiotic activity of acidic sophorolipids with potential for facilitating wound healing［J］. Antimicrob Agents Chemother， 2017， 61（5）： E02547.
17	SAERENS K M J， SAEY L， SOETAERT W. One-step production of unacetylated sophorolipids by an acetyltransferase negative Candida bombicola［J］. Biotechnol Bioeng， 2011， 108（12）： 2923-2931.
18	VAN B， BUYST D， MARTINS J C， et al. Synthesis of bolaform biosurfactants by an engineered starmerella bombicola yeast［J］. Biotechnol Bioeng， 2016， 113（12）： 2644-2651.
19	马孝萌. 去乙酰化槐糖脂基因工程菌株的构建及发酵条件优化［D］. 青岛：中国海洋大学， 2023.
	MA X M. Construction of genetically engineered strains producing deacetylated sophorolipids and its optimization of fermentation conditions［D］. Qingdao： Ocean University of China， 2023.
20	宋丹丹，梁生康，王江涛. 槐糖脂生物表面活性剂的结构特征及理化性质初探［J］. 环境化学， 2011， 30（8）： 1474-1479.
	SONG D D， LIANG S K， WANG J T. Structural characteristics and physical-chemical properties of sophorolipid biosurfactants［J］. Environ Chem， 2011， 30（8）： 1474-1479.
21	孙旭璐，陈慧，郑伟. 改性大豆磷脂的HLB值检测方法优化研究［J］. 粮食与油脂， 2015， 28（4）： 47-48.
	SUN X L， CHEN H， ZHENG W. Study on the optimal method for detecting of modified soybean phospholipids HLB value‍［J］. Cereals Oils， 2015， 28（4）： 47-48.
22	周家华，崔英德. 表面活性剂HLB值的分析测定与计算Ⅰ∶ HLB值的分析测定［J］. 精细石油化工， 2001（2）： 11-14.
	ZHOU J H， CUI Y D. Measurement and calculation of HLB value of surfactants： the measurement of HLB value［J］. Spec Petrochem， 2001（2）： 11-14.
23	DAVID J T， ROBERT L R. Commercial synthesis of monoalkly phosphates‍［J］. J Surfactants Deterg， 2002， 5（2）： 169-172.
24	李双阳，王佳颖，郭高强，等. BOLA型表面活性剂的研究进展［J］. 广州化工， 2015， 43（5）： 18-20.
	LI S Y， WANG J Y， GUO G Q， et al. Progress in research work with respect to bolaform surfactants‍［J］. Guangzhou Chem Ind， 2015， 43（5）： 18-20.
25	周家华，崔英德，吴雅红. 表面活性剂HLB值的分析测定与计算 Ⅱ.HLB值的计算［J］. 精细石油化工， 2001（4）： 38-41.
	ZHOU J H， CUI Y D， WU Y H. Measurement and calculation of HLB value of surfactants： the calculation of HLB value‍［J］. Spec Petrochem， 2001（4）： 38-41.
26	KRAYNIK A M. The structure of random foam［J］. Adv Eng Mater， 2006， 8（9）： 900-906.
27	KUMAR V， PAL N， JANGIR A K， et al. Dynamic interfacial properties and tuning aqueous foamability stabilized by cationic surfactants in terms of their structural hydrophobicity， free drainage and bubble extent‍［J］. Colloid Surface A， 2020， 588： 124362.
28	WANG C， FANG H， GONG Q，et al. Roles of catanionic surfactant mixtures on the stability of foams in the presence of oil‍［J］. Energ Fuel， 2016， 30（8）： 6355-6364.
29	赵修太，王泉，王增宝，等. 高温高盐油藏常用起泡剂研究进展［J］. 材料导报， 2016， 30（5）： 75-80.
	ZHAO X T， WANG Q， WANG Z B， et al. Commonly used foaming agents in high temperature and high salinity reservoir［J］. Mater Rep， 2016， 30（5）： 75-80.
30	SUN L， SUN X H， ZHANG Y C， et al. Stability of high-salinity-enhanced foam： surface behavior and thin-film drainage‍［J］. Petrol Sci， 2023， 20（4）： 2343-2353.
31	刘晓敏. 驱油用泡沫体系研究［D］. 无锡：江南大学， 2023.
	LIU X M. Study on foam systems for enhanced oil recovery［D］. Wuxi： Jiangnan University， 2023.
32	胡贝贝，袁悦，周小平，等. 基于丝氨酸的Bola型表面活性剂的合成及理化性质［J］. 沈阳药科大学学报， 2016， 33（6）： 419-425.
	HU B B， YUAN Y， ZHOU X P， et al. Synthesis of a novel Bola amphiphile based on serine and investigation of its physicochemical properties［J］. J Shenyang Pharm Univ， 2016， 33（6）： 419-425.
33	VAN RENTERGHEM L， ROELANTS S， BACCILE N， et al. From lab to market： an integrated bioprocess design approach for new-to-nature biosurfactants produced by Starmerella bombicola‍［J］. Biotechnol Bioeng， 2018， 115（5）： 1195-1206.

	Type		Molecular formula	m/z of ［M+H］⁺		Full name of component	w/%
	Fatty acid groups	L/A/B^*	Molecular formula	Measured value	Simulation value	Full name of component	w/%
Nonacetylated	C18∶2	A	C₃₀H₅₂O₁₃	621.342 1	621.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid	0.32
Monoacetylated	C16∶0	L	C₃₀H₅₂O₁₃	621.743 5	621.340 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1′，4"- lactone-6"-monoacetyl ester	0.11
	C18∶0	L	C₃₂H₅₆O₁₃	649.369 7	649.372 2	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanoic acid-1′，4"- lactone-6"-monoacetyl ester	0.18
	C18∶1	L	C₃₂H₅₄O₁₃	647.358 1	647.356 5	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1′，4"-lactone-6"- monoacetyl ester	5.29
	C18∶2	L	C₃₂H₅₂O₁₃	645.341 9	645.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1′，4"-lactone-6"-monoacetyl ester	1.60
Diacetylated	C16∶0	L	C₃₂H₅₄O₁₄	663.360 4	663.351 4	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1′，4"- lactone-6′，6"-diacetyl ester	12.27
	C16∶1	L	C₃₂H₅₂O₁₄	661.338 6	661.335 7	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecenoic acid-1′，4"-lactone-6'，6"-diacetyl ester	0.15
	C18∶0	L	C₃₄H₅₈O₁₄	691.384 9	691.382 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanoic acid-1′，4"-lactone-6′，6"-diacetyl ester	16.97
	C18∶1	L	C₃₄H₅₆O₁₄	689.375 4	689.367 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1′，4"-lactone-6′，6"-diacetyl ester	42.47
	C18∶2	L	C₃₄H₅₄O₁₄	687.356 3	687.351 4	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1′，4"-lactone-6'，6"-diacetyl ester	18.82
	C16∶2	A	C₃₂H₅₂O₁₅	677.331 9	677.330 6	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanodienoic acid-6′，6″-diacetyl ester	1.29
	C18∶1	A	C₃₄H₅₈O₁₅	707.382 5	707.377 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-6′，6″-diacetylester	0.19
	C18∶2	A	C₃₄H₅₆O₁₅	705.351 6	705.362 0	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-6′，6″- diacetyl ester	0.18
	C22∶1	A	C₃₈H₆₆O₁₅	745.432 9	745.429 9	21-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］- docosenoic acid -6′，6″- diacetyl ester	0.16

	Type		Molecular formula	m/z of ［M+H］⁺		Full name of component	w/%
	Fatty acid groups	L/A/B^*	Molecular formula	Measured value	Simulation value	Full name of component	w/%
Nonacetylated	C18∶2	A	C₃₀H₅₂O₁₃	621.342 1	621.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid	0.32
Monoacetylated	C16∶0	L	C₃₀H₅₂O₁₃	621.743 5	621.340 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1′，4"- lactone-6"-monoacetyl ester	0.11
	C18∶0	L	C₃₂H₅₆O₁₃	649.369 7	649.372 2	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanoic acid-1′，4"- lactone-6"-monoacetyl ester	0.18
	C18∶1	L	C₃₂H₅₄O₁₃	647.358 1	647.356 5	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1′，4"-lactone-6"- monoacetyl ester	5.29
	C18∶2	L	C₃₂H₅₂O₁₃	645.341 9	645.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1′，4"-lactone-6"-monoacetyl ester	1.60
Diacetylated	C16∶0	L	C₃₂H₅₄O₁₄	663.360 4	663.351 4	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1′，4"- lactone-6′，6"-diacetyl ester	12.27
	C16∶1	L	C₃₂H₅₂O₁₄	661.338 6	661.335 7	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecenoic acid-1′，4"-lactone-6'，6"-diacetyl ester	0.15
	C18∶0	L	C₃₄H₅₈O₁₄	691.384 9	691.382 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanoic acid-1′，4"-lactone-6′，6"-diacetyl ester	16.97
	C18∶1	L	C₃₄H₅₆O₁₄	689.375 4	689.367 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1′，4"-lactone-6′，6"-diacetyl ester	42.47
	C18∶2	L	C₃₄H₅₄O₁₄	687.356 3	687.351 4	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1′，4"-lactone-6'，6"-diacetyl ester	18.82
	C16∶2	A	C₃₂H₅₂O₁₅	677.331 9	677.330 6	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanodienoic acid-6′，6″-diacetyl ester	1.29
	C18∶1	A	C₃₄H₅₈O₁₅	707.382 5	707.377 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-6′，6″-diacetylester	0.19
	C18∶2	A	C₃₄H₅₆O₁₅	705.351 6	705.362 0	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-6′，6″- diacetyl ester	0.18
	C22∶1	A	C₃₈H₆₆O₁₅	745.432 9	745.429 9	21-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］- docosenoic acid -6′，6″- diacetyl ester	0.16

	Type		Molecular formula	m/z of ［M+H］⁺		Full name of component	w/%
	Fatty acid groups	L/A/B^*	Molecular formula	Measured value	Simulation value	Full name of component	w/%
Nonacetylated	C16∶2	L	C₂₈H₄₆O₁₂	575.310 4	575.298 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecanodienoic acid-1′，4″-lactone	8.73
	C18∶1	L	C₃₀H₅₂O₁₂	605.354 5	605.345 9	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-oleic acid-1′，4″-lactone	16.50
	C18∶3	L	C₃₀H₄₈O₁₂	637.365 2	637.335 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］- octadecanotrienoic acid-1′，4″-lactone	1.76
	C16∶0	A	C₂₈H₅₂O₁₃	597.509 0	597.340 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecenoic acid	16.96
	C16∶0	A	C₂₈H₅₂O₁₃	597.347 9	597.340 8	16-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecenoic acid	1.74
	C18∶0	A	C₃₀H₅₆O₁₃	625.386 3	625.372 2	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-octadecanoic acid	6.37
	C18∶1	A	C₃₀H₅₄O₁₃	623.364 2	623.356 5	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-oleic acid	10.18
	C18∶2	A	C₃₀H₅₂O₁₃	621.356 7	621.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-octadecanodienoic acid	1.38
	C20∶1	A	C₃₂H₅₆O₁₂	651.395 8	651.387 9	19-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-eicosenoic acid	3.59
	C24∶0	A	C₃₆H₇₄O₁₂	709.496 9	709.466 4	23-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-lignoceric acid	1.49
	C16∶0	B	C₄₀H₇₂O₂₃	921.450 2	921.446 4	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	6.60
	C16∶1	B	C₄₀H₇₀O₂₃	901.435 5	919.430 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］- hexadecenoic acid -1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	10.16
	C18∶1	B	C₄₂H₇₄O₂₃	947.469 3	947.462 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	2.74
	C18∶2	B	C₄₂H₇₂O₂₃	945.485 5	945.446 4	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	2.36
Monoacetylated	C18∶1	A	C₃₂H₅₆O₁₄	669.365 0	665.367 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-6″-monoacetyl ester	8.27
	C18∶1	B	C₄₄H₇₆O₂₃	989.478 5	989.472 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester-6″-monoacetyl ester	0.36
	C26∶0	B	C₅₂H₉₄O₂₃	1103.664 2	1103.614 0	25-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexacosanoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester-6″-monoacetyl ester	0.81

	Type		Molecular formula	m/z of ［M+H］⁺		Full name of component	w/%
	Fatty acid groups	L/A/B^*	Molecular formula	Measured value	Simulation value	Full name of component	w/%
Nonacetylated	C16∶2	L	C₂₈H₄₆O₁₂	575.310 4	575.298 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecanodienoic acid-1′，4″-lactone	8.73
	C18∶1	L	C₃₀H₅₂O₁₂	605.354 5	605.345 9	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-oleic acid-1′，4″-lactone	16.50
	C18∶3	L	C₃₀H₄₈O₁₂	637.365 2	637.335 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］- octadecanotrienoic acid-1′，4″-lactone	1.76
	C16∶0	A	C₂₈H₅₂O₁₃	597.509 0	597.340 8	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecenoic acid	16.96
	C16∶0	A	C₂₈H₅₂O₁₃	597.347 9	597.340 8	16-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-hexadecenoic acid	1.74
	C18∶0	A	C₃₀H₅₆O₁₃	625.386 3	625.372 2	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-octadecanoic acid	6.37
	C18∶1	A	C₃₀H₅₄O₁₃	623.364 2	623.356 5	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-oleic acid	10.18
	C18∶2	A	C₃₀H₅₂O₁₃	621.356 7	621.340 8	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-octadecanodienoic acid	1.38
	C20∶1	A	C₃₂H₅₆O₁₂	651.395 8	651.387 9	19-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-eicosenoic acid	3.59
	C24∶0	A	C₃₆H₇₄O₁₂	709.496 9	709.466 4	23-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）- O-］-lignoceric acid	1.49
	C16∶0	B	C₄₀H₇₂O₂₃	921.450 2	921.446 4	15-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexadecanoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	6.60
	C16∶1	B	C₄₀H₇₀O₂₃	901.435 5	919.430 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］- hexadecenoic acid -1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	10.16
	C18∶1	B	C₄₂H₇₄O₂₃	947.469 3	947.462 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	2.74
	C18∶2	B	C₄₂H₇₂O₂₃	945.485 5	945.446 4	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-octadecanodienoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester	2.36
Monoacetylated	C18∶1	A	C₃₂H₅₆O₁₄	669.365 0	665.367 1	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-6″-monoacetyl ester	8.27
	C18∶1	B	C₄₄H₇₆O₂₃	989.478 5	989.472 7	17-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-oleic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester-6″-monoacetyl ester	0.36
	C26∶0	B	C₅₂H₉₄O₂₃	1103.664 2	1103.614 0	25-L-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-hexacosanoic acid-1-［（2′-O-β-D-glucopyranosyl-β-D-glucopyranosyl）-O-］-ester-6″-monoacetyl ester	0.81

	Acetylated SLs	Nonacetylated SLs
Solubility/（g·L^-1）	33.8	485.8
Cloud point/℃	36	Not detected
Acid resistance	Conditional acid resistance	Non acid resistance
Alkali resistance	Non alkali resistance	Non alkali resistance
Hard water resistance/grade	5	5