水果口感细腻柔软，富含维生素、矿物质、膳食纤维等营养素，深受人们喜爱。我国水果资源丰富，但具有明显的时令性，多集中在6月～9月，受环境及自身生理等影响，多数水果保质期较短，且采后贮藏技术相对缺乏，导致鲜果极易腐败，降低了水果的品质[1]；此外，在水果生长过程伴随有较多的次果、落果、坏果等副产物[2]。因此，关于水果的精深加工一直是食品领域关注的热点之一。
随着人们生活水平提高和健康意识增强，水果现有产品形式过于单一，已无法满足市场需求，因此迫切需要创新产品形式。基于益生菌发酵技术研发的新型发酵果汁，营养价值高，且具有独特风味[3]。发酵果汁以新鲜水果为原料，通过菌种发酵而得。目前的发酵果汁主要有两种，一种是单一发酵果汁，如发酵苹果汁[4]、发酵梨汁[5]、发酵葡萄汁[6]、发酵蓝莓汁[7]等；另一种是混合发酵果汁，如苹果、柚子和梨的混合发酵果汁[8]；所用菌种也可分两类，一是单一菌种，如植物乳杆菌发酵猕猴桃汁[9]；二是混合菌种，如保加利亚乳杆菌与嗜热链球菌混合发酵枸杞果汁[10]。果汁经发酵后会生成大量乳酸、氨基酸、短链脂肪酸等营养物质，提高水果的营养功效，已成为现代食品发展的一个新趋势。本文将主要阐述发酵果汁的菌种选择、发酵工艺及营养价值等方面的研究进展，并讨论其发展前景，旨在为发酵果汁的研发提供参考。
1 发酵果汁的菌种
益生菌（probiotics）具有调节肠道微生态平衡、维持心血管健康、提高免疫力等功效[11]，其种类丰富，目前广泛应用的主要有以下几类：1）乳杆菌类，如植物乳杆菌、嗜酸乳杆菌、干酪乳杆菌等；2）双歧杆菌类，如嗜热双歧杆菌、长双歧杆菌、短双歧杆菌等；3）其他菌属，如嗜热链球菌、酵母菌、醋酸菌等。随着益生菌在水果加工中的深入研究与广泛应用，果汁逐渐成为益生菌发展的潜力载体[12]。
1.1 乳酸菌及其在果汁中的应用
乳酸菌（lactic acid bacteria，LAB）在自然界分布广泛，其对机体的益生作用已有较多报道。经乳酸菌发酵处理不仅可以增强食品风味，还会生成具有特殊营养功效的物质，如糖苷酶[13]，以及一些具有抑菌作用的物质，如有机酸、细菌素等[14]，显著提高食品品质。
发酵过程中乳酸菌的代谢作用使果汁中的酚类物质增加，从而提高果汁的抗氧化性[15]。张宏志等[16]向蓝莓黑莓混合果汁中接种植物乳杆菌、乳酸乳杆菌和肠膜明串珠杆菌进行发酵，发酵后果汁的pH值、总糖浓度下降，总酸、总酚含量升高，抗氧化性增强。王茹等[17]以肠膜明串珠菌和植物乳杆菌混合发酵枸杞汁，经工艺优化所得产品乳酸菌含量达2.1×108CFU/mL，且具有较强的抑菌效果。乳酸菌发酵果汁可增强机体免疫功能，提高免疫力。尹曼等[18]研究了以乳酸菌发酵的复合果蔬汁提取物对小鼠脾淋巴细胞增殖的影响，发现复合发酵果蔬汁可促进T/B淋巴细胞增殖，各提取物均有较好的保护脾淋巴细胞免受氧化应激损伤的作用，证明复合发酵果蔬汁具有免疫调节的功效。Dimitrovski等[19]以植物乳杆菌发酵菊芋汁，经发酵的菊芋汁活菌数、乳酸含量较高，且发现发酵菊芋汁与蓝莓汁混合可提高产品货架期。DI CAGNO等[20]以植物乳杆菌发酵石榴汁，发酵后醇类、酮类、烯烃类、萜类、苯类化合物含量显著增加，而醛类和硫化物含量减少，表明乳酸菌发酵可提升石榴汁的感官品质。由此可见，乳酸菌发酵果汁不仅口感风味较好，还能提高果汁的品质。
1.2 酵母菌及其在果汁中的应用
酵母菌（Saccharomyces）具有较为广泛的应用价值，利用酵母菌发酵果汁，能适当降低糖含量，提高酚类、黄酮以及维生素C等活性抗氧化物质的含量[21]。酵母菌在食品行业已有较多应用，如啤酒、奶酪等的制作[22-23]。近年来，酵母菌在发酵果汁中也有研究应用。高淑芳等[24]以酵母菌发酵火龙果汁，发现所得产品具有火龙果香和发酵香气。刘景等[25]从自然发酵果蔬液中分离出酵母菌，接种于苹果汁中，显著改善了原苹果汁的香味。邹颖等[26]利用干酪乳杆菌与三种酵母菌发酵荔枝汁，结果表明复合发酵能促进干酪乳杆菌的生长，且发酵后的荔枝汁糖度下降，具有较好的抗氧化效果。Fracassetti等[27]利用酵母菌发酵制备了一种葡萄和猕猴桃混合果汁。由此可知，酵母菌能在果汁中稳定生长繁殖，且可改善果汁品质，已成功应用于果汁发酵。
1.3 醋酸菌及其在果汁中的应用
醋酸菌（Acetobacter aceti）是一类革兰氏阴性细菌，广泛分布于自然界[28]，其发酵过程产生的醋酸，具有清理肠胃、促进消化吸收营养物质的功效。尹爱国等[29]以基酒为基础，在最优醋酸发酵工艺条件下制得的龙眼和柠檬复合果汁的总酸为68 g/L，相较于原果汁，其醇类、酯类物质含量增加，具有独特风味。张艺等[30]以油橄榄果汁为原料，研究酿造食醋的液态发酵工艺，通过最佳发酵工艺得到总酸为51.2 g/L的油橄榄风味食醋，相较于相同工艺下生产的白醋，其含有更多的黄酮、多酚等抗氧化成分。然而，目前关于醋酸菌发酵果汁的报道还较少，有待深入研究。
2 发酵果汁的工艺
不同的水果基质具有迥异的理化性质，导致发酵工艺的多样，主要是由于不同菌种在不同果汁中的生长情况与活性不同；且对于同种果汁而言，单一菌种、混合菌种和不同复配比的混合菌种也会影响其发酵后的品质，如总多酚、总黄酮、总糖等含量，以及自由基清除率、风味、色泽等[26，31-32]。几种常见发酵果汁的工艺条件如表1所示。
表1 几种发酵果汁的工艺条件
Table 1 Technological conditions of several fermented fruit juice
原料 菌种 工艺条件 产品品质酸浆果汁[33]嗜酸乳杆菌∶植物乳杆菌=2∶1（质量比）活菌数1.86×109CFU/mL，乳酸含量32 g/L，口感顺滑，外观均匀陈皮柑汁[34] 植物乳杆菌 0.8%β-环糊精，45℃保温90 min脱苦，2%接种量，10%木糖醇添加量，26℃发酵7 h，后转0℃～4℃低温熟化24 h 0.01%β-环糊精，0.04%阿斯巴甜，0.02%安赛蜜，8%果葡糖浆，0.03%柠檬酸，10%接种量，37℃恒温发酵20 h 8%果糖、3%酵母浸粉；7%接种量，75%果汁浓度，28℃发酵34 h 活菌数1.0×108CFU/mL，乳酸含量17.93 g/L，口感润滑，有沙果香与发酵香苹果草莓复合果汁[36]活菌数1.43×108CFU/mL，质地均匀，口感柔和，有陈皮柑香气沙果汁[35] 保加利亚乳杆菌∶植物乳杆菌=1∶2（质量比）嗜酸乳杆菌 苹果汁∶草莓汁=1∶1（质量比），1%接种量，37℃发酵40 h 可溶性固形物含量18.5%，糖度15°Bx，pH值4.23猕猴桃汁[37]植物乳杆菌∶嗜酸乳杆菌=1∶1（质量比）3%接种量，36℃发酵24 h 活菌数2.19×108CFU/mL，有猕猴桃果香与乳酸风味沙棘汁[38] 酵母菌 0.5%果胶酶，45℃酶解2 h，0.03%酵母，28℃发酵24 h，8%白砂糖，0.1%苹果酸，0.1%单甘脂，0.1%黄原胶，0.05%羧甲基纤维素钠口感圆润柔和，酸甜适中，呈橙黄色，有沙棘果香和发酵香气
3 发酵果汁的营养功效
发酵果汁能最大限度地保留水果的营养成分，且发酵过程会产生较多的营养物质，提高果汁品质的同时，还能改善肠道环境，提高免疫力[39]。Nayak等[40]研究发酵诺丽果汁对糖尿病小鼠的作用效果，发现其具有降血糖和保护肝脏作用。
发酵过程微生物的代谢作用会使果汁中的酚类含量、酶系组成等发生变化，进而改变果汁品质，如风味、自由基清除率、抗氧化性等[26]，发酵给果汁带来的主要物质变化如图1所示[41-46]。
图1 发酵果汁的营养物质变化
Fig.1 Changes of nutrients in fermented juice
有研究发现，经乳酸菌发酵后，果汁中总酚、黄酮、维生素、矿物质、胞外多糖等含量增加[47]。卢嘉懿[48]研究了5种不同果蔬汁发酵前后的物质变化，发现挥发性风味物质含量及种类、芳香族氨基酸、苯衍生风味物质均增加，醛类物质和游离氨基酸减少，表明发酵过程游离氨基酸可能分解生成了一些小分子挥发性呈味物质。刘秋豆等[49]研究发现经乳酸菌发酵的芒果汁，总酚含量增加，抗氧化活性提高，果汁品质明显提升。Lu等[50]分别以瑞士乳杆菌、副干酪乳杆菌和鼠李糖乳杆菌发酵杨桃汁，结果发现杨桃汁中的醛类和酯类等异味物质含量显著下降，而酮类、醇类和脂肪酸含量在一定程度上增加，改善了杨桃汁的香味。Park等[51]发现经乳酸菌发酵后混合浆果汁的抗氧化活性显著提高，且新增了34种挥发性化合物。
发酵果汁的营养价值具体如表2所示。
表2 发酵果汁的营养功效分析
Table 2 Analysis of nutritional efficacy of fermented fruit juice
营养功效 作用机制调节肠道菌群平衡 乳酸、醋酸等有机酸可降低肠道pH值，抑制病原菌[52-54]降低脂肪和胆固醇 有机酸和氨基酸促进脂肪代谢，降解胆固醇，尼克酸和维生素促使胆固醇排出[55-56]软化血管、降血压 三萜类物质和黄酮促进心血管扩张、增加冠状动脉血流量[57-58]消除疲劳、提高免疫 有机酸和氨基酸促进有氧代谢，清除乳酸；通过激活免疫细胞、促进单核细胞吞噬能力和调控免疫细胞因子分泌来调节免疫[59-60]促进消化吸收、润肠通便有机酸促进肠道蠕动、胃液分泌；多种消化酶促进消化[61-62]养颜美容、延缓衰老 含抗氧化物质，防止形成过氧化物，延缓细胞衰老[34，63]
4 国内外发酵果汁的发展现状与趋势
益生菌发酵果汁已经成为国内研究热点，郑欣[55]以荔枝为原料，研究干酪乳杆菌、保加利亚乳杆菌和肠膜状明串珠菌发酵荔枝汁的工艺，解决了荔枝含糖量高、果香不突出的问题，开发出新型的荔枝汁发酵饮料。李海萍[64]研究乳酸菌发酵柚子汁的工艺，脱去了柚子汁的苦味，得到口感较好的柚子发酵果汁。范俊华等[8]以植物乳杆菌发酵柚子、苹果、梨的混合果汁，研制出有柔和发酵风味与浓郁果香的发酵复合果汁。杨冲等[65]利用乳酸菌发酵南丰蜜桔汁，发酵后的果汁总酸度升高，多酚和总黄酮含量显著增加，表明乳酸菌发酵可改善原果汁风味，提高其营养价值和保健功效。
国外也有大量关于益生菌发酵果汁的研究。Mantzourani等[66]研究植物乳杆菌发酵对石榴汁品质的影响，结果表明发酵石榴汁比未发酵石榴汁含有更多挥发性化合物（醇类、酮类和酯类）；在贮藏期间，发酵石榴汁的总酚含量与抗氧化活性均高于未发酵石榴汁。Guo等[67]研究发现鲜诺丽果汁的体外抗氧化能力虽高于发酵诺丽果汁，但发酵果汁对小鼠急性酒精肝损伤有更好的预防效果，表明体外和体内的抗氧化效果和机制有差异。RICCI等[68]研究植物乳杆菌、干酪乳杆菌、副干酪乳杆菌和鼠李糖乳杆菌在樱桃汁中的生长能力，结果表明经发酵后蔗糖和苹果酸含量明显降低，乳酸和挥发性物质含量增加；且植物乳杆菌能调节果汁的酚类成分，增加二氢咖啡酸（抗氧化物质）和苯乳酸（抑菌物质）的含量，鼠李糖乳杆菌和副干酪乳杆菌可增加芳香化合物的含量，为发酵樱桃汁菌种选择提供了参考。Muhialdin等[69]以植物乳杆菌发酵火龙果汁，发现发酵前后果汁的抗氧化活性无明显差异，但发酵后的果汁对大肠杆菌、伤寒沙门氏菌、铜绿假单胞菌和金黄色葡萄球菌的抑菌活性较高，且赖氨酸、丙氨酸、乙酸、丁二酸、乳酸、葡萄糖、异丁酸和甜菜碱的含量增加，证明乳酸菌发酵可改善和提高果汁的品质。
5 结论与展望
本文介绍了发酵果汁的研究进展，并对其菌种选择、工艺条件、营养功效进行归纳和分析。目前，针对发酵果汁的报道还较少，市场上的益生菌发酵饮品仍以发酵乳制品为主[70]。利用益生菌发酵果汁，既可保留水果原有营养成分，也能增加水果的营养价值，提高果品品质。此外，以非乳原料作为益生菌的发酵基质，可以为乳糖不耐受人群提供摄入益生菌的新选择。因此，在当下消费者追求科学平衡的膳食模式大背景下，代表健康、营养、绿色的发酵果汁将是未来食品行业的发展方向和研究热点，具有巨大的商业化潜力，有望成为大众青睐的新型功能性饮料。
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