谷朊粉加工产生的大量高浓度蛋白质废水，富含多种有机污染物，直接排放会导致水体富营养化，引发藻类大量繁殖，破坏水生生态平衡，还会造成蛋白质资源浪费。因此，开展谷朊粉废水蛋白质回收研究具有重要的环境与资源意义。

本研究基于不同方法的原理及蛋白质回收应用潜力，系统评估了碱提酸沉法、超声辅助碱提酸沉法、天然有机高分子絮凝法、微生物絮凝剂法以及综合法的效果，从蛋白质回收率、结构特性以及废水净化程度，为废水资源化和环境治理提供理论及技术参考。

（1）碱提酸沉法与超声辅助碱提酸沉法效果探究：通过析因实验、最陡爬坡实验和响应面优化实验，确定碱提酸沉法最佳条件为碱提pH 9.6、温度45℃、时间55 min，酸沉pH 6、温度30℃、时间20 min，实际回收率达（37.94±2.52）%。在上述基础上引入超声辅助，最佳条件为超声功率140 W、时间7 min，回收率提升至（53.83±1.37）%，且可提升必需氨基酸回收。此外，对碱提酸沉回收处理后的水样进行水质指标检测，检测结果显示，该法对COD、TP、TN和NH₃-N的去除率分别为67.09%、22.99%、20.66%和19.86%，表明该方法在一定程度上能够有效净化谷朊粉废水，但回收率有待提升，故而探究天然有机高分子絮凝法。

（2）天然有机高分子絮凝法效果探究：从壳聚糖、卡拉胶和海藻酸钠中筛选出海藻酸钠，其蛋白回收效果最佳。结果表明，海藻酸钠在pH 3、剂量1.0 g/L、絮凝温度25℃，回收率高达（99.29±0.36）%。其回收蛋白质的氨基酸保留情况较好，部分氨基酸含量高于谷朊粉。此外，对COD、TP、TN和NH₃-N的去除率分别为86.28%、2.31%、53.45%、26.09%。尽管海藻酸钠在蛋白质回收方面优势明显，但考虑到废水处理的复杂性，接着探讨微生物絮凝法。

（3）微生物絮凝法效果探究：从谷朊粉废水中纯化、筛选出絮凝率达（77.54±1.44）%的塔宾曲霉（B菌株）。经培养优化，B菌发酵液冻干处理后，20 mg/mL冻干粉复溶液对高岭土悬浊液絮凝率达87.91%。通过单因素及正交实验，确定该方法的最佳条件为pH为9、冻干粉复溶液浓度20 mg/mL、静置时间为150 min，此时，蛋白回收率达到（47.26±2.11）%。同时，微生物絮凝法对COD、TP、TN和NH₃-N的去除率分别为65.68%、6.20%、20.35%、11.64%。该方法基于微生物独特代谢机制，为谷朊粉废水处理提供新思路，但其综合效果有待提升，进而研究综合法。

（4）综合法效果探究：碱提酸沉法和微生物絮凝分别作为预处理与超滤结合。实验结果显示，两种预处理结合超滤后的蛋白回收率分别为85.84%和93.22%；对应的超滤滤液COD、TP和TN均展现良好的去除效果，去除率均达到90%以上，而碱提酸沉预处理下的超滤滤液的NH₃-N去除率为80.83%，略高于微生物絮凝预处理下的70.07%。综合法在蛋白质回收与废水净化多方面取得较好的综合效果。但面临膜污染与成本问题。

综上，本研究的多种蛋白质回收方法各有优劣，未来需进一步优化以克服局限。

The large amount of high-concentration protein wastewater generated from gluten powder processing is rich in various organic pollutants. Direct discharge of this wastewater can lead to eutrophication of water bodies, triggering the massive reproduction of algae, disrupting the aquatic ecological balance, and also causing waste of protein resources. Therefore, conducting research on the recovery of proteins from gluten powder wastewater is of great environmental and resource significance.

Based on the principles of different methods and their potential applications in protein recovery, this study systematically evaluated the effects of the alkali - extraction and acid - precipitation method, the ultrasonic - assisted alkali - extraction and acid - precipitation method, the natural organic polymer flocculation method, the microbial flocculant method, and the comprehensive method. From the aspects of protein recovery rate, structural characteristics, and the degree of wastewater purification, this study provides insights for wastewater resource utilization and environmental governance.

1. Investigation on the Effects of Alkaline Extraction - Acid Precipitation Method and Ultrasound - Assisted Alkaline Extraction - Acid Precipitation Method

Through factorial experiments, steepest ascent experiments, and response surface optimization experiments, the optimal conditions for the alkali - extraction and acid - precipitation method were determined as follows: the pH value for alkali - extraction was 9.6, the temperature was 45°C, the time was 55 min; the pH value for acid - precipitation was 6, the temperature was 30°C, and the time was 20 min. The actual recovery rate reached (37.94±2.52)%. On this basis, when ultrasonic assistance was introduced, the optimal conditions were an ultrasonic power of 140 W and a time of 7 min, and the recovery rate increased to (53.83±1.37)%, and the recovery of essential amino acids could also be improved. In addition, the water quality indicators of the water samples recovered by the alkali - extraction and acid - precipitation method were detected. The detection results showed that the removal rates of COD, TP, TN, and NH₃-N by this method were 67.09%, 22.99%, 20.66%, and 19.86% respectively. This indicates that this method can effectively purify gluten powder wastewater to a certain extent, but the recovery rate needs to be improved. Therefore, the natural organic polymer flocculation method was explored.

2. Investigation on the Effect of Natural Organic Polymer Flocculation Method

Among chitosan, carrageenan, and sodium alginate, sodium alginate was screened out as having the best protein recovery effect. The results showed that when the pH value was 3, the dosage was 1.0 g/L, and the flocculation temperature was 25°C, the recovery rate of sodium alginate was as high as (99.29±0.36)%. The amino acid retention of the recovered protein was good, and the content of some amino acids was higher than that of gluten powder. In addition, the removal rates of COD, TP, TN, and NH₃-N were 86.28%, 2.31%, 53.45%, and 26.09% respectively. Although sodium alginate has obvious advantages in protein recovery, considering the complexity of wastewater treatment, the microbial flocculation method was then explored.

3. Investigation on the Effect of Microbial Flocculation Method

Aspergillus tubingensis (strain B) with a flocculation rate of (77.54±1.44)% was screened out from the concentrated gluten powder wastewater. Through single - factor experiments, the main factors affecting the protein recovery rate were initially explored, and then orthogonal experiments were designed to comprehensively analyze the interaction of these factors. As a result, the optimal conditions of this method were determined as a pH value of 9, a freeze - dried powder re - solution concentration of 20 mg/mL, and a flocculation time of 150 min. At this time, the recovery rate reached (47.26±2.11)%. At the same time, the removal rates of COD, TP, TN, and NH₃-N by the microbial flocculation method were 65.68%, 6.20%, 20.35%, and 11.64% respectively. This method provides a new idea for the treatment of gluten powder wastewater based on the unique metabolic mechanism of microorganisms, but its comprehensive effect needs to be improved. Therefore, the comprehensive method was further studied.

4. Investigation on the Effect of the Comprehensive Method

The alkali - extraction and acid - precipitation method and microbial flocculation were selected as pretreatment methods respectively to be combined with ultrafiltration due to their unique advantages in initial protein separation and wastewater pre - treatment. The experimental results showed that the protein recovery rates after ultrafiltration with the two pretreatments were 85.84% and 93.22% respectively; the ultrafiltration filtrates had good removal effects on COD, TP, and TN, with the removal rates all reaching more than 90%. The removal rate of NH₃-N in the ultrafiltration filtrate under the alkali - extraction and acid - precipitation pretreatment was 80.83%, slightly higher than 70.07% under the microbial flocculation pretreatment. The comprehensive method achieved good comprehensive effects in many aspects such as protein recovery and wastewater purification. However, it faces problems such as membrane fouling and cost.In conclusion, the various protein recovery methods investigated in this study each present distinct advantages and limitations. Future research should focus on further optimization to overcome these constraints.

In conclusion, the various protein recovery methods in this study have their own advantages and disadvantages, and further optimization is needed in the future to overcome the limitations.

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